Differential expression of PACAP receptors in postnatal rat brain

Pituitary Adenylate Cyclase Activating Polypeptide (PACAP) is a multi-functional neuropeptide that acts through activation of three common G-protein coupled receptors (VPAC1, VPAC2 and PAC1). In this study, we have investigated the gene expression profile of PAC1 isoforms (Hop1, Hip, Hip-Hop) and VPAC1, VPAC2 receptors in distinct brain regions during different stages of rat postnatal development. Using quantitative real time PCR approach we found that PAC1 isoforms were highly expressed in the cortex of newborns with marked decrease in expression during later stages of development. In contrast, mRNA levels of VPAC1, VPAC2 receptors were markedly lower in newborns in comparison to later developmental stages. Expression of PAC1 isoforms predominated in the hippocampus, while expression of VPAC1 was more prominent in the cortex and VPAC2 in the striatum and hippocampus. In addition we found that during early stages of postnatal development the expression of PAC1 receptor in the hippocampus was significantly higher in females than in males. No sex dependent differences in expression were observed for the VPAC1 and VPAC2 receptors. In summary, differential expression of PAC1, VPAC1 and VPAC2 receptors during postnatal development as well as gender dependent differences of PAC1 receptor expression in the hippocampus, will contribute to our understanding of the role of PACAP/VIP signaling system in normal brain development and function.     

Differential anatomical and cellular patterns of connexin43 expression during postnatal development of rat brain.

We have shown previously that connexin43 in the adult rat central nervous system (CNS) is predominantly localized at astrocytic gap junctions. Here we document immunohistochemically the emergence of connexin43-immunoreactive (connexin43-IR) structures and the regional patterns of connexin43 expression during postnatal maturation of the rat brain. On Western blots, connexin43 was detected in brain samples at postnatal day (P) 5, the earliest age studied. Immunohistochemically, most brain regions displayed a characteristic sequence of transient immunoreactive profiles that ultimately gave rise to the uneven distribution of the protein seen in adults. Generally, brains at P1-P5 exhibited long, fibrous connexin43-IR elements which were identified as radial glial cells. This fibrous immunostaining was considerably diminished at P5 and was replaced by short immunoreactive processes which predominated up to P10. These processes had a stellate appearance, emanated from partially stained astrocytic cell bodies and were heterogeneously distributed throughout the developing brain. By P15, there occurred only punctate immunolabelling similar to that seen in adult brain. Some brain regions including the amygdaloid complex, septohypothalamic nucleus, preoptic hypothalamus, zona incerta, ependyma and subfornical organ were exceptional in that they displayed adult immunostaining patterns at early postnatal ages suggesting a precocious maturation of gap junctions in these areas. We conclude that the highly heterogeneous distribution of connexin43-immunoreactivity among defined nuclear structures in adult brain does not reflect an antecedent requirement for connexin43 in early brain morphogenesis, but rather is related to the development of neuronal activity, the establishment of functional circuitry and the contribution of astrocytic gap junctions to glial metabolic coupling and potassium spatial buffering in the mature CNS.     

Carboxypeptidase E (enkephalin convertase): mRNA distribution in rat brain by in situ hybridization

Carboxypeptidase E (CPE), also referred to as enkephalin convertase or carboxypeptidase H (EC 3.4.17.10), is present in neurotransmitter secretory granules and can remove C-terminal basic residues following endopeptidase cleavage during peptide processing. Using in situ hybridization with 35S-labeled oligonucleotide probes, we have mapped the localization of CPE mRNA in the rat brain. Specificity for CPE was confirmed by control experiments, which included production of identical patterns hybridization with 3 different antisense oligonucleotide probes, loss of label with RNase pretreatment of sections or co-incubation with excess unlabeled probe, and lack of labeling with sense orientation probes. In addition, the regional distribution of CPE mRNA by Northern blot analysis corresponded with distribution of labeling by in situ hybridization. The highest levels of CPE mRNA were found to be present in the pyramidal cells of the hippocampus, the pituitary anterior and intermediate lobes, the ependymal cells of the lateral ventricle, the endopiriform nucleus, the basolateral amygdala, the supraoptic nucleus, and the paraventricular nucleus. Intermediate levels were present in the thalamus, medial geniculate nucleus, lateral septal nucleus, piriform and entorhinal cortex, nucleus of the tractus solitarius, cerebellar cortex, pontine nuclei, and inferior olive. The lowest levels were found in the hippocampal granule cell layer, lateral hypothalamus, globus pallidus, and brain stem reticular formation. Ibotenic acid lesions of the hippocampus eliminated the majority of the label, which had been present over pyramidal cells, though labeling was increased over areas of reactive gliosis, suggesting that activated astrocytes can also synthesize CPE mRNA. In general, the localization of CPE mRNA in the rat brain corresponded to the distribution of enkephalin and other peptide neurotransmitter-synthesizing neurons, though CPE mRNA was also present in neurons that do not secrete known peptides and in reactive glia. The widespread yet specific localization of CPE mRNA in the rat brain suggests it may be an excellent marker for peptide synthesizing cells in the CNS.     

Differential expression of peroxisomal matrix and membrane proteins during postnatal development of mouse brain

In peroxisomal biogenesis disorders, serious neurological abnormalities can be observed in the patients and the respective knockout mouse models. As a prerequisite for a better understanding of the relationship between the absence of peroxisomes and the observed neuropathology, knowledge of the regional and cell-type specific distribution of peroxisomal proteins in mouse brain is necessary. Therefore, we investigated the expression of distinct peroxins, peroxisomal membrane and matrix proteins (e.g. Pex5p, Pex14p, Pex13p, PMP70, catalase, peroxisomal thiolase, Acox1, "SKL"-PTS1 proteins) by indirect immunofluorescence 1) in primary cultures of the medial neocortex, hippocampus, and cerebellum of newborn mice and 2) in paraffin sections of mouse brain of different ages (newborn-adult). Quantitative analysis revealed a comparable abundance (number/microm(2)) of peroxisomes in cultured neurons and astrocytes of all three brain regions. In contrast, catalase immunoreactivity was higher in cultured astrocytes than in neurons. In mouse brain tissue, the abundance of peroxisomes decreased by half during postnatal development, also exhibiting prominent differences between distinct brain regions and cell types. Catalase protein levels in neuronal peroxisomes, however, decreased much more strongly in the neocortex, CA1-3 areas of the hippocampus, dentate gyrus, cerebellar nuclei, and cerebellar cortex but remained high in Bergmann glia and other astrocytes, epithelial cells of the choroid plexus, and ependyma. Similar age-dependent changes were found for thiolase and Acox1 protein levels. Developmental changes were confirmed by Western blot analysis using enriched peroxisomal and cytosolic fractions of the brain tissue as well as by measurement of catalase activity.     

Effects of prenatal hypoxia on expression of thioredoxin-1 in the rat hippocampus at different stages of postnatal ontogeny

We studied the expression of the antioxidant protein thioredoxin-1 in the hippocampus of rat pups that were subjected to hypoxia at prenatal days 14–16, at the 3rd and 14th days after their birth, and at 80–90 days when the pups became mature. In spite of the specific features of certain hippocampal regions and the presence of exceptions related to them, 3 days after birth we observed a general trend to a decrease in thioredoxin-1 expression in rats subjected to hypoxia, as compared to the control animals of the same age. Then, thioredoxin-1 expression was substantially increased on the 14th day of postnatal life. In adult rats, thioredoxin-1 expression decreased again. These changes were very prominent in the CA1 field of the hippocampus. A decrease in the activity of the physiological antioxidant system is an important index of cell-membrane homeostasis, whose impairment under hypoxic conditions can result in the development of neurodegenerative diseases.     

Spatial relationship between NSCs/NPCs and microvessels in rat brain along prenatal and postnatal development

Neurogenesis and angiogenesis are two parallel processes that occur in brain development and repair, and so share some molecular signals. In order to better understand the interaction between the genesis of neural cells and vessels during brain development, the density of microvessels and the number of nestin positive neural stem/neural progenitor cells (NSCs/NPCs) around microvasculature in various brain regions was quantified. Results showed that the density of microvessels remained at a relative low level during embryonic development and dramatically increased after postnatal day 3 (P3), especially in subventricular zone. The number of nestin positive NSCs/NPCs per microvessel in neurogenic brain regions continually increased with fetal brain development and then gradually dropped down during postnatal development. The highest density of NSCs/NPCs appeared at postnatal day 1 (P1) and dramatically decreased after P3. Similar pattern was observed in striatum. In the olfactory bulb, the cerebral cortex and cerebellum, the dramatic decrease of NSCs/NPCs density appeared after P7, especially in the cerebral cortex. Our results demonstrated that anatomically, the spatial relationship between NSCs/NPCs and microvessels changed during brain development. The alteration patterns in neurogenic brain regions differed from non-neurogenic brain regions.     

Differential ontogeny of three putative catecholamine cell types in the postnatal rat retina

The development of tyrosine hydroxylase (TH) and phenylethanolamine N-methyltransferase (PNMT) in the rat retina was investigated using the indirect immunofluorescence technique. Two types of TH-positive cells were found. The first appeared at postnatal day 2, in the vitreal half of the inner nuclear layer (INL). Single fibres from these neurones, bifurcating in and innervating layer 3 of the inner plexiform layer (IPL) were seen at day 4. This first type of TH-positive cell was most numerous at day 15, but thereafter disappeared before adulthood. At day 5, a more intensely staining TH-immunoreactive neurone became visible, occupying a more proximal part of the INL, and projecting multiple fibres to layer 1 of the IPL. In contrast, PNMT-positive cells, in the vitreal half of the INL and in the ganglion cell layer (GCL), sending single varicose axons to layer 3 of the IPL, were first apparent only at day 10, achieving a disposition similar to that of the adult by days 15-16 postnatal. Analysis of adjacent sections stained with antibodies to TH and PNMT revealed that neither type of TH-positive neurone also contained PNMT-like immunoreactivity. It is concluded that although both of the rate-limiting enzymes of the catecholamine synthetic pathway are present in the developing rat retina, they occur in 3 mutually exclusive populations of neurones.     

GABA-stimulated chloride uptake and enhancement by diazepam in synaptoneurosomes from rat brain during prenatal and postnatal development

The present study was undertaken to evaluate the ability of diazepam (DZ) to elicit a response in the brains of developing animals. gamma-Aminobutyric acid (GABA)-stimulated 36Cl- uptake in the presence and absence of DZ was measured in synaptoneurosomal preparations from whole brain of fetal rats at 20 and 21 days gestation and from cerebral cortex of rats at 7, 14, 21, 28 and 60-90 days postnatal age. The ability of GABA to stimulate 36Cl- uptake in a concentration-dependent manner was evident from gestational day 20. The EC50 for GABA stimulation but not the maximum stimulation increased significantly from day 20 to day 21 of gestation. Postnatally, only moderate changes in the EC50 were evident, but the maximum 36Cl- transported increased significantly from 7 to 14 days and remained stable thereafter. DZ enhanced GABA-mediated 36Cl- influx from 20 days gestation, and this enhancement was seen as a decrease in the EC50 for GABA stimulation. However, DZ also significantly increased maximum GABA-stimulated 36Cl- transport in synaptoneurosomal preparations at 21 days gestation and at 7 days postnatal age, a response to DZ not seen at older ages. DZ had a less robust effect on GABA-mediated 36Cl- transport at 28 days postnatal age than at any other age, a result consistent with functional observations of decreased responsivity to DZ in late juvenile, early adolescent rats. The benzodiazepine antagonist Ro 15-1788 prevented the effect of DA on GABA-stimulated 36Cl- uptake in tissue from 21-day fetuses and from 60- to 90-day-old adults.(ABSTRACT TRUNCATED AT 250 WORDS)     

Serotoninergic development in the postnatal rat brain

Summary Various characteristics of the developing serotoninergic system in the brain of rats aged 1 to 28 days were studied biochemically.The levels of the precursor amino acid tryptophan showed a maximal increase in the blood, brain and cerebrospinal fluid (CSF) during the 7th and 10th postnatal days. The development of tryptophan hydroxylase activity measuredin vivo by means of 5-hydroxytryptophan (5-HTP) accumulation after NSD 1015 was closely related to the 5-hydroxytryptamine (5-HT) levels at the various ages. 5-HTP accumulation and 5-HT levels increased most markedly after the second postnatal week. 5-Hydroxyindoleacetic acid (5-HIAA) levels were found to increase rapidly in the brain but somewhat more slowly in the CSF during the second week of postnatal development. Regional studies of 5-HTP accumulation after NSD 1015, 5-HT and 5-HIAA levels indicated a caudal to rostral way of maturation.The disappearance of 5-HT was measured after inhibition of tryptophan hydroxylase with H 22/54. The half-life generally decreased in the various brain parts with advancing age, and in the younger animals the shortest half-life was found in the most caudal brain parts. At 28 days of age the half-life was similar in all brain parts studied. These results indicate the existence of an adult like nerve impulse flow in the 5-HT neurons in the brain stem region of the newborn rats. The results from this investigation clearly indicate that the maturation of the different biochemical parameters of the 5-HT pathways develop in a caudal to rostral direction.The study also supports the view that tryptophan hydroxylase may be the limiting step in the development of the serotoninergic system.     

Differential biosynthesis and posttranslational processing of vasopressin and oxytocin in rat brain during embryonic and postnatal development

The biosynthesis and posttranslational processing of arginine vasopressin (AVP) and oxytocin (OT) peptides in the developing rat brain and pituitary were studied using antibodies and complementary separation methods that permitted a quantitative radioimmunoassay (RIA) analysis of precursor, intermediate, and completely processed forms of the peptides. Precursor forms of the peptides were first detected in rat brain as early as embryonic day (E) 15 for AVP and E17 for OT. Proteolytic cleavage products of the precursors were detected 1 d later for both peptides. AVP was present in a fully processed (amidated) from immediately (E16) and throughout fetal development. OT was cleaved from its precursor starting on E18 but remained in an intermediate (C-terminal extended) form until E21, when amidated OT was first detected in the pituitary. Hence, Pro-AVP processing in the fetus was immediate and complete, whereas Pro-OT processing in the fetus was much slower and incomplete, resulting in the generation of partially processed, nonamidated stable forms of the peptide (OT-Gly10, OT-Gly10-Lys11, and OT-Gly10-Lys11-Arg12). The presence of OT-Gly10-Lys11-Arg12 as a major, stable intermediate form, indicated that the in vivo pattern of endoproteolytic cleavage occurred principally at the C-terminus of the pair of basic amino acids at the tripeptide spacer sequence (Gly-Lys-Arg) in the precursor. Although both precursors were first expressed nearly simultaneously in the brain, the steady-state levels of the precursors were very different throughout fetal life. From E16-E21, the quantities of AVP precursors and peptides were 5- to 10-fold greater than those of OT, suggesting a much higher level of precursor biosynthesis in the AVP neurons. In addition to these differences in the regulation of biosynthesis and processing, AVP peptides were axonally transported to the pituitary 3 d earlier than OT peptides, and in far greater (20-fold) abundance. The early presence and abundance of amidated AVP in the brain and pituitary suggests a trophic function for this peptide during development.     

Angiotensin II receptor subtypes and angiotensin-converting enzyme in the fetal rat brain

Angiotensin II (ANG II) receptor subtypes (AT₁, displaced by losartan, and AT₂, displaced by CGP 42112A) were characterized by quantitative autoraography after incubation with the ANG II agonist [¹²⁵I]Sar¹-ANG II, in specific brain nuclei of 19-day-old rat embryos. Binding to AT₁ receptors, located in the subfornical organ, paraventricular nucleus, nucleus of the solitary tract and choroid plexus, was sensitive to incubation with GTPγS. The sensitivity of AT₂ receptors to GTPγS was heterogeneous. In the ventral thalamic, rostral hypoglossal and medial geniculate nuclei, and in the locus coeruleus, binding to AT₂ receptors was sensitive to GTPγS and these areas belong to the AT_2A subgroup. Conversely, in the inferior olive, medial (fastigial) cerebellar nucleus and caudal part of the hypoglossal nucleus, areas belonging to the AT_2B subgroup, binding was insensitive to GTPγS. AT₂ receptors were also present in cerebral arteries. In the fetal anterior pituitary, AT₁ receptors predominated. The angiotensin-converting enzyme (ACE; EC 3.4.15.1) was studied by autoradiography with the selective inhibitor [¹²⁵I]351A. In 19-day-old embryos, ACE was highly expressed in chroid plexus, with high concentrations in subfornical organ, posterior pituitary and cerebral arteries. No ACE binding was detected in extrapyramidal structures or anterior pituitary in 19-day-old embryos.     

Differential expression of the estrogen receptors alpha and beta during postnatal development of the rat cerebellum

Estrogen receptor (ER) beta is a dominant ER subtype in the adult cerebellum. However, it is not known if this is also the case for the developing cerebellum. In the present study, quantitative real-time RT-PCR demonstrated that levels of cerebellar ERalpha mRNA in neonatal pups were significantly higher than in adults. In contrast, expression levels of cerebellar ERbeta mRNA remained significantly unchanged during postnatal development. In situ hybridization and immunohistochemistry demonstrated that ERalpha mRNA and protein were predominantly expressed by Purkinje cells at all ages examined. ERalpha-expressing Purkinje cells were confined to the anterior lobes at postnatal day 7 (P7) but distributed in most lobes at P14 and P21. In the adult cerebellum, however, only a few ERalpha-immunoreactive Purkinje cells were observed. Thus, ERalpha expression was transiently increased during the time when Purkinje cell dendritic growth and synapse formation proceed, suggesting that a role for ERalpha in Purkinje cell differentiation. ERbeta expression occurred in Golgi type neurons in the granular layer at P7, Purkinje cells at P14, and basket cells in the molecular layer at P21 and was detected in all the cell types in the adult cerebellum, suggesting a role for ERbeta associated with neuronal differentiation and maintenance. Furthermore, double-labeled immunofluorescence for ERalpha and ERbeta demonstrated their colocalization in Purkinje cells at P14, suggesting a possibility of their interaction. The discrete expression profiles for ERalpha and ERbeta in the developing cerebellum suggest the two ERs play distinct roles in cerebellar development.     

Differential sensitivity of tachykinin vs. enkephalin gene expression in the posterior striatum in response to acute p-chloroamphetamine treatment during postnatal development

The acute effects of the monoamine releaser p-chloroamphetamine (pCA, 10 mg/kg, i.p.) on preprotachykinin (PPT) and preproenkephalin (PPE) mRNA expression in the anterior (A-STR) vs. posterior (P-STR) striatum were studied in rodents at postnatal days (PND) 10, 21 and 35. Northern analysis 4 h post-injection yielded no significant mRNA changes within the A-STR of any pCA group. However, significant increases (80–200% of saline control) in PPT mRNA levels occurred within the P-STR at all three postnatal ages. Interestingly, pCA did not increase PPE mRNA levels within the P-STR until PND 35 (150% of saline control). Such observation suggests that tachykinin neurons of the P-STR achieve an earlier monoamine-responsive signal transduction linkage to gene regulation as compared to enkephalin neurons. Given its predominance in the caudal regions of the striatum, 5-HT neurotransmission at the 5-HT₂ receptor is suggested to play a central role in this mechanism.     

Early postnatal development of rat brain: in vivo diffusion tensor imaging

Perinatal hypoxia is a major cause of neurodevelopmental deficits. Neuronal migration patterns are particularly sensitive to perinatal hypoxia/ischemia and are associated with the clinical deficits. The rat model of hypoxia/ischemia at P7 mimics that of perinatal injury in humans. Before assessing the effects of postnatal injury on brain development, it is essential to determine the normal developmental trajectories of various brain structures in individual animals. In vivo longitudinal diffusion tensor imaging (DTI) was performed from postnatal day 0 (P0) to P56 on Wistar rats. The DTI metrics, mean diffusivity (MD), fractional anisotropy (FA), axial (lambdal) and radial (lambdat) diffusivities, were determined for four gray matter and eight white matter structures. The FA of the cortical plate and the body of corpus callosum decreased significantly during the first 3 weeks after birth. The decrease in the cortical plate's FA value was associated mainly with an increase in lambdat. The initial decrease in FA of corpus callosum was associated with a significant decrease in lambdal. The FA of corpus callosum increased during the rest of the observational period, which was mainly associated with a decrease in lambdat. The FA of gray matter structures, hippocampus, caudate putamen, and cortical mantle did not show significant changes between P0 and P56. In contrast, the majority of white matter structures showed significant changes between P0 and P56. These temporal changes in the DTI metrics were related to the neuronal and axonal pruning and myelination that are known to occur in the developing brain.     

The ontogeny and localization of glutamine synthetase gene expression in rat brain

A 2.4 kb cDNA clone containing the coding sequence for glutamine synthetase (GS) was isolated from a rat brain cDNA library, and a probe constructed from this cDNA was utilized in Northern analysis of total RNA to study the tissue distribution and the ontogeny of GS mRNA expression in the rat brain from embryonic day 14 (E14) to adulthood. The levels of GS mRNA were highest in the brain, followed by kidney and liver. In the brain, the GS message was detected as early as E14, earlier than it can be detected by either enzymatic assays of GS activity or by immunocytochemical localization of GS. The relatively low levels of GS mRNA seen at E14 increase to a peak around the time of birth, and in the second postnatal week another rise in GS message occurs approaching adult levels by P15. Localization of GS to astrocytes in the brain was confirmed using both immunocytochemistry and in situ hybridization.     

Differential regulation of the low-affinity nerve growth factor receptor during postnatal development of the rat brain.

We studied the temporal and spatial localization of the low-affinity nerve growth factor receptor (LNGF-R) during the early postnatal period in rat brain in order to understand better the relationship between nerve growth factor (NGF)-like responsiveness and the development of specific central neuronal populations. Four different developmental patterns of LNGF-R mRNA hybridization were found in this study. First, some neurons contain high levels of LNGF-R mRNA from postnatal time points into adulthood, as exemplified by neurons of the cholinergic basal forebrain and mesencephalic trigeminal nucleus. Second, several cell groups exhibit robust hybridization during the early postnatal period but contain much reduced levels of LNGF-R mRNA in the adult brain. These include striatal neurons, Purkinje cells of the cerebellum, and several medullary nuclei. A third group of cells produces the LNGF-R transiently during development, including cranial nerve nuclei of the brainstem, the periolivary nuclei complex, the reticular formation, and the deep cerebellar nuclei. Finally, cell populations which may exist only transiently during central nervous system (CNS) development, such as subplate neurons of the cerebral cortex, appear to express the LNGF-R during only a brief period. These results show that the LNGF-R gene is differentially regulated in a cell type-specific manner during development, and suggests that diverse neuronal populations require only transient growth factor sensitivity, while others exhibit NGF-like responsitivity into maturity.     

Differential expression and localization of the phosphorylated and nonphosphorylated neurofilaments during the early postnatal development of rat hippocampus

Neurofilament (NF) proteins are expressed in most mature neurons in the central nervous system. Although they play a crucial role in neuronal growth, organization, shape, and plasticity, their expression pattern and cellular distribution in the developing hippocampus remain unknown. In the present study, we have used Western blotting and immunocytochemistry to study the low- (NF-L), medium- (NF-M), and high- (NF-H) molecular-weight NF proteins; phosphorylated epitopes of NF-M and NF-H; and a nonphosphorylated epitope of NF-H in the early postnatal (through P1-P21) development of the rat hippocampus. During the first postnatal week, NF-M was the most abundantly expressed NF, followed by NF-L, whereas the expression of NF-H was very low. Through P7-P14, the expression of NF-H increased dramatically and later began to plateau, as also occurred in the expression of NF-M and NF-L. At P1, no NF-M immunopositive cell bodies were detected, but cell processes in the CA1-CA3 fields were faintly immunopositive for NF-M and for the phosphorylated epitopes of NF-M and NF-H. At P7, CA3 pyramidal neurons were strongly immunopositive for NF-L and NF-H, but not for NF-M. The axons of granule cells, the mossy fibers (MFs), were NF-L and NF-M positive through P7-P21 but were NF-H immunonegative at all ages. Although they stained strongly for the phosphorylated NF-M and NF-H at P7, the staining intensity sharply decreased at P14 and remained so at P21. The cell bodies of CA1 pyramidal neurons and granule cells remained immunonegative against all five antibodies in all age groups. Our results show a different time course in the expression and differential cell type and cellular localization of the NF proteins in the developing hippocampus. These developmental changes could be of importance in determining the reactivity of hippocampal neurons in pathological conditions in the immature hippocampus.     

Differential expression of FGF-2 isoforms in the rat adrenal medulla during postnatal development in vivo

Basic fibroblast growth factor (FGF-2) isoforms of the adrenal medulla are differentially expressed during rat postnatal development. While the 18 and 23 kDa isoforms continuously rise towards the adult expression level, the 21 kDa isoform displays a peak expression at postnatal day 28. The peak expression of the 21 kDa isoform correlates with the peak of the corticosterone concentration during postnatal development. Together with the previously demonstrated increase of the 21 kDa isoform in the adrenal medulla in vivo after glucocorticoid administration these results suggest that the differential regulation of the FGF-2 isoforms could be a physiologically occurring mechanism.