Neural damage following retrochiasmatic knife cuts in the rat

In order to define neuronal damage produced by sham, frontal and frontolateral hypothalamic knife cuts known to effect neuroendocrine and behavioral variables, we designed a retractable knife capable of delivering horseradish peroxidase as a cut was being produced. Damaged neurons filled with homogeneous reaction product were found in virtually all hypothalamic nuclei. A similar distribution of damage was produced by frontolateral (FLC), frontal (FC) or sham cuts—the primary differences being the greater density of damaged neurons in hypothalamic nuclei and labeling of the ventral tegmental area of Tsai in FLC and FC animals. The difference between FC and FLC animals was primarily one of degree of damage. The amount of damage produced by the sham surgery is consistent with the idea that sham surgeries produce endocrine dysfunctions intermediate between frontal cut animals and unoperated controls. It is interesting to note that a number of the nuclei damaged by these surgeries provide aminergic innervation to the hypothalamus.     

Afferent connections to the lateral hypothalamus: A horseradish peroxidase study in the rat

Horseradish peroxidase, 13% Sigma Type VI, was administered iontophoretically to the mid lateral hypothalamus (LH) of male hooded rats. Animals were perfused intracardially on the following day and brains were removed and sliced in the coronal or sagittal planes into 30–50 μm sections. Sections were processed with DAB and BDH for the brown and blue reaction products and later examined by bright and dark field microscopy for the presence and location of retrogradely labeled neurons. Results indicate that a significant number of afferent connections to the LH originate in the olfactory and accumbens nuclei, pyriform cortex, olfactory tracts, magnocellular and medial preoptic and anterior hypothalamic regions, stria terminalis, stria hypothalamic tract, diagonal tract of Broca, caudate-putamen and globus pallidus, internal capsule, lateral septal nuclei, lateral preoptic area and anterior medial forebrain bundle, the various amygdaloid nuclei, zona incerta, perifornical region, dorsal and ventral medial hypothalamic areas, supraoptic, paraventricular and periventricular nuclei, posterior hypothalamus and medial forebrain bundle, ventral thalamic nuclei, the fields of Forel, arcuate and mammillary nuclei, adjacent to the fasciculus retroflexus, in the ventral tegmental area of Tsai, interpeduncular nucleus, substantia nigra, mesencephalic reticular formation, periaqueductal gray, locus coeruleus and parabrachial region. Results are discussed in terms of previous anatomical and neurophysiological data, probable pathways, and the function of LH neurons.     

A horseradish peroxidase study of the cerebellorubral pathway in the rat

The distribution and character of labeled neurons within the cerebellar nuclei of the rat were studied after injection of horseradish peroxidase into the red nucleus. Cells possessing horseradish peroxidase activity were found throughout the contralateral nucleus lateralis (dentate) and interposed complex and were absent from the nucleus medialis (fastigial). Relatively more labeled neurons were present within the anterior part of the interposed complex than in its posterior portion. Peroxidase-stained cells were characteristically large and multipolar, whereas small neurons throughout these centers were devoid of activity. An ipsilateral projection to the red nucleus was not observed.     

Trigeminal nerve and temporomandibular joint of the cat: A horseradish peroxidase study

The mesencephalic nucleus of the trigeminal nerve is considered to contain the cell bodies of the first-order neurons that have peripheral connections in the temporomandibular joint capsule. Through use of retrograde transport of horseradish peroxidase, this concept is challenged. The results indicate that a relatively specific region of the mandibular division of the trigeminal ganglion of the cat contains the first-order neurons innervating the temporomandibular joint capsule.     

A horseradish peroxidase study of feline mandibular incisors

A 15% horseradish peroxidase solution (0.5 μl) was placed in cavity preparations of ipsilateral central (7), first (2), or second (2) mandibular incisors of adult cats. Following a 24-h survival period, the cats were sacrificed and their trigeminal ganglia removed for histochemical processing. The results suggest that there is no peripheral cross-innervation of feline mandibular incisors, which supports previous studies demonstrating ipsilaterality of feline canine teeth.     

Mesencephalic and pontine afferent fiber system to the facial neucleus in the cat: a study using the horseradish peroxidase and silver impregnation techniques.

The mesencephalic and pontine afferent fiber system to the medial or lateral part of the facial nucleus was examined in the cat by the horseradish peroxidase and the Fink-Heimer methods. The medial part of the facial nucleus was observed to receive afferent fibers mainly from the perioculomotor and the midbrain paralemniscal regions. The perioculomotor region, which included the oculomotor nucleus, the Edinger-Westphal nucleus, the interstitial nucleus of Cajal, the nucleus of Darkschewitsch, and the ventral part of the periaqueducatal gray, sent fibers to the facial nucleus bilaterally with a slight dominance of the ipsilateral distribution. The midbrain paralemniscal region, which was just medial to the dorsal part of the medial lemniscus at caudal levels of the superior colliculus, sent numerous fibers to the contralateral facial nucleus. In contrast, the lateral part of the facial nucleus received many afferent fibers from the ventral part of the parabrachial nuclei; the parabrachiofacial fibers were distributed bilaterally with a marked predominance of the ipsilateral distribution. The existence of crossed rubrofacial fibers was confirmed. Some neurons in the ventral part of the nucleus pontis centralis oralis were found to send fibers to the facial nucleus bilaterally with a predominance of the contralateral distribution.     

Cortical projections to the superior colliculus in the macaque monkey: A retrograde study using horseradish peroxidase

The topical and laminar distribution of corticotectal cells, as well as their size and morphology, were studied in the macaque monkey with the horseradish peroxidase (HRP) technique. After HRP injections restricted primarily to the superficial layers of the colliculus, labelled cells were found in visual cortex (areas 17, 18, and 19) and both in the frontal eye field (area 8) and the adjacent part of premotor cortex (area 6). The clustering of labelled cells in visual cortex indicated that each of the anatomically and functionally distinct visual areas has its own set of collicular projections. When intermediate and deeper layers of the colliculus were injected, labelled cells were found also in posterior parietal cortex (area 7) where they were concentrated mainly on the posterior bank of the intraparietal fissure, in inferotemporal cortex (areas 20 and 21), in auditory cortex (area 22), in the somatosensory representation SII (anterior bank of sylvian fissure, area 2), in upper insular cortex (area 14), in motor cortex (area 4), in premotor cortex (area 6), and in prefrontal cortex (area 9). In the motor and premotor cortex, labelled cells formed a continuous band which appeared to stretch across finger-hand-arm-shoulder-neck representation. Similarly, the cluster of labelled cells in area 2 may correspond to the finger-hand representation of SII. The cortical regions not containing labelled cells were the somatosensory representation SI (areas 3, 1 and 2) and the infraorbital cortex. Labelled cells were restricted to layer V of all cortical areas except in the primary visual cortex, where labelled cells were found in both layer V and layer VI. The size spectrum of corticotectal cells ranged from 14.8 μm (average diameter) in area 17 to 27.8 μm in area 6, comprising cells as small as 8 μm and as large as 45 μm. Labelled cells in posterior parietal (area 7), in auditory (area 22), and in motor cortex (area 4) were small and distributed over only a narrow range of sizes. Those in premotor cortex (area 6) were often large and had a wide range in size distribution. The differences in size and morphology of corticotectal neurons suggest that they do not form a uniform class of neurons.     

Spinal afferents to lamina IX of the cervical enlargement in the rat studied by the retrograde transport of horseradish peroxidase

Spinal neurons that project to the ventrolateral, dorsolateral and ventromedial portions of lamina IX of the cervical enlargement in the rat were investigated by means of horseradish peroxidase retrograde transport. In the cervical and upper thoracic segments, labeled neurons were observed ipsilaterally in laminae V–VIII and contralaterally in laminae VII–VIII. In the lower thoracic and upper lumbar segments, labeled neurons were seen after HRP injection into the ventrolateral part of lamina IX, and were distributed mainly in the lateral parts of the ipsilateral laminae V–VI.     

Differences in horseradish peroxidase labeling of sensory, motor and sympathetic neurons following chronic axotomy of the rat sural nerve

In an attempt to clarify the ultimate fate of permanently axotomized adult primary neurons, horseradish peroxidase (HRP) was used as a cell marker to label the motor, sensory and postganglionic sympathetic neurons of rat sural nerves which had been sectioned at the ankle and prevented from regenerating for periods of up to 80 weeks. Axotomy did not affect sympathetic neurons, but resulted 4 weeks later in a sudden reduction in the number of labeled sensory and motor cells which persisted to the end of the study. The missing neuronal population amounted to 44.4% and 45.9% respectively of the normal sensory and motor contingent and included most of the large afferent and efferent neurons. However, examination of sural nerves at the thigh, 30 mm proximal to the neuroma, revealed marked axonal atrophy but no change in the number of myelinated and unmyelinated fibers up to 52 weeks after axotomy. Such prolonged survival of the peripheral processes is indirect evidence that axotomized neurons can endure long-term detachment from their end organs and suggests that the lack of HRP labeling in certain sensory and motor neurons does not imply their degeneration, but expresses one of many retrograde dysfunctions triggered by axotomy.     

Disruption of maternal behaviour in virgin and postparturient rats following sagittal plane knife cuts in the preoptic area-hypothalamus

Parasagittal knife cuts along the medial preoptic area-medial anterior hypothalamus were placed at either the medial (near lateral or NL cuts) or lateral (far lateral or FL cuts) border of the medial forebrain bundle in separate groups of virgin (Experiment 1) and postparturient (Experiment 2) rats. FL cuts were placed so as to spare preoptic-hypothalamic efferent connections with the medial forebrain bundle. NL and FL knife cuts were equally effective in preventing the induction of maternal behaviour produced by repeated exposure of virgin rats to foster pups. Both types of cuts also reduced nest building in virgins. In postparturient rats, NL and FL cuts both abolished pup retrieval and reduced nursing behaviour. However, only NL cuts disrupted lactation and nest building. NL cuts also produced chronic hyperthermia. The deficits in maternal behaviour and nest building may be independent of this hyperthermia, as shaving the fur in animals with NL cuts reduced body temperatures to control levels but did not restore these behaviours. These findings indicate that while preoptic/hypothalamic connections through the medial forebrain bundle are important for nest building and possibly lactation, other lateral connections must also be important for pup retrieval and nursing behaviour. The identity of these connections remains to be determined.     

The hypothalamic ‘tuberoinfundibular’ system of the rat as demonstrated by horseradish peroxidase (HRP) microiontophoresis

Microiontophoresis of horseradish peroxidase (20%) into the median eminence of the rat has allowed visualization of perikarya and axon projections of the tuberoinfundubular system after retrograde transport. Cells projecting to the median eminence were found in the periventricular regions of the hypothalamus and were particularly pronounced in dorsal portions of the rostral arcuate nucleus, the medial division of the paraventricular nucleus, and within the anterior periventricular nucleus. Labeling of perikarya within the ventromedial nucleus was rarely found. No labeling by HRP was found within cells of the dorsomedial, anterior, suprachiasmatic, preoptic, lateral hypothalamic nuclei or within the septal and amygdaloid nuclei. Axons from identifiable cells were located within the periventricular neuropil and contained within the baso-lateral portions of the hypothalamic-hypophysial tract.     

Labelling of olfactory bulb glomeruli following horseradish peroxidase lavage of the nasal cavity

The transport of horseradish peroxidase (HRP) from the nasal cavity to the olfactory bulb was examined in rat. HRP was present primarily in the olfactory nerve and glomerular layer. In some animals the glomeruli were densely filled with product while in others there was considerable interglomerular variation in density. Examination of the decalcified noses revealed a restricted distribution of HRP in those rats with partially labelled olfactory bulbs. The presence of small groups of densely labelled glomeruli was also noted using the 2-deoxyglucose method to examine odor-induced metabolic activity.     

A direct projection from the hypothalamus to the area postrema in the rat, as demonstrated by the HRP and autoradiographic methods

A direct hypothalamic afferent to the area postrema was found in the rat. After an injection of tritiated amino acids into the hypothalamus, labeled fibers and terminals were observed in the entire part of the area postrema, with dense patches of silver grains present in places. Cells of origin of this novel projection were identified with the retrograde horseradish peroxidase (HRP) method in the dorsal hypothalamic area at the most rostral level of the dorsomedial hypothalamic nucleus.     

Parabrachial nucleus neurons projecting to the lower brain stem and the spinal cord. A study in the cat by the Fink-Heimer and the horseradish peroxidase methods

Direct projections from the parabrachial nucleus (PBN) to the lower brain stem and the spinal cord were examined in the cat by the Fink-Heimer and the horseradish peroxidase (HRP) methods. After placing lesions in the PBN, many fine degenerated fibers were seen contralaterally in the ventromedial portions of the caudal pontine reticular formation, and ipsilaterally in the lateral portions of the facial nucleus, the regions around the hypoglossal nucleus, and the regions around the ambiguus nucleus; some degenerated fibers were traced ipsilaterally down to the spinal cord. Subsequently, HRP injections were attempted into these regions where many fine degenerated fibers were observed. In cats injected with HRP into the lateral portions of the facial nucleus, the regions around the hypoglossal nucleus, the regions around the ambiguus nucleus, or the first cervical cord segment, many HRP-labeled neurons were seen in the ventral portions of the PBN. The mean of the average soma diameters of the PBN neurons labeled with HRP injected into the regions around the hypoglossal nucleus or the first cervical cord segment was significantly larger than that of the PBN neurons labeled with HRP injected into the lateral portions of the facial nucleus or the regions around the ambiguus nucleus.     

Serotonergic projections to the caudal brain stem: a double label study using horseradish peroxidase and serotonin immunocytochemistry

Cells of origin of serotonergic and non-serotonergic projections to the caudal brain stem in the primate were examined using a double label technique. Following HRP injections into medullary raphe nuclei and the adjacent reticular formation double labeled cells were found in the dorsal raphe nucleus, the central superior nucleus and the ventrolateral tegmentum. Retrogradely labeled cells that did not stain for serotonin-like immunoreactivity were found primarily in the periaqueductal gray (PAG) and the mesencephalic and pontine reticular formation. The results are discussed in relation to the descending pathway(s) mediating the effects of PAG stimulation.     

The location of the preganglionic neurons that innervate the submandibular gland of the cat. A horseradish peroxidase study

Horseradish peroxidase was injected under pressure in the main excretory duct of the submandibular gland of cats, to locate the perikarya of the preganglionic parasympathetic salivatory neurons in the brainstem. Labeled multipolar neurons were found ipsilaterally in the lateral reticular formation, where they extended from the rostral plane that contains the genu of the facial nerve to the caudal plane that contains the rostral pole of the dorsal motor nucleus of the vagus nerve.     

A direct hypothalamic projection to the superior salivatory nucleus neurons in the rat. A study using anterograde autoradiographic and retrograde HRP methods

The location of the superior salivatory nucleus and terminal labelings of the hypothalamic descending fibers were demonstrated in the nucleus reticularis parvocellularis using HRP and the autoradiographic techniques, respectively. When both techniques were used in the same animals, some HRP-labeled neurons were seen among the accumulations of silver grains, suggesting pericellular terminations. The present study demonstrates that the hypothalamic efferents project directly to the superior salivatory nucleus innervating salivary and lacrimal glands.     

A new device for making multiple injections of small volumes of horseradish peroxidase into the spinal cord

A device which allows multiple injections of horseradish peroxidase to be made into the spinal cord with minimal tissue damage is described. The unit is constructed from commercially available capillary glass, silicon vacuum grease and fast drying dental cement. It is cheap and simple to construct and permits the injection of known volumes as small as 7.8 nl.     

Preganglionic innervation of the adrenal gland of the rat using horseradish peroxidase

Using the tetramethyl benzidine method for the demonstration of horseradish peroxidase, it was possible to label retrogradely spinal neurons whose axons project to the adrenal medulla in both young and adult rats. In both age groups, labeled neurons were found in the ipsilateral intermediolateral cell column of the T2 through T13 segments of the spinal cord, with most of the labeling in the T6 to T10 segments. The neurons labeled with horseradish peroxidase also demonstrated a strong acetylcholinesterase activity.