The origins of innervation of the canine caudal pharyngeal muscles: an HRP study.

In deglutition, movements of the tongue and oropharynx direct a bolus to the laryngopharynx. The major muscles of this region, which includes the 'cricopharyngeal sphincter', must undergo sequential relaxation and contraction for correct swallowing action. The innervation of the caudal pharyngeal muscles involved in this action in the dog have not been determined previously by sensitive neuroanatomical techniques. In this study, the location of efferent and afferent neurons innervating the left cricopharyngeus, thyropharyngeus and hyopharyngeus muscles was determined by horseradish peroxidase (HRP) histochemistry in 7 puppies. Labeled cells were found ipsilaterally in the supraspinal nucleus, nucleus ambiguus (including nucleus retrofacialis) and nucleus intercalatus of all animals, in the parasympathetic nucleus of X (dorsal vagal efferent nucleus) of 6 animals, and in the hypoglossal nucleus of 4 animals. Small numbers of HRP-labeled cells were found contralaterally in the supraspinal nucleus of all animals, and in the rostral nucleus ambiguus, in the nucleus intercalatus and the parasympathetic nucleus of X of fewer animals. This defines a more extensive source of efferent neurons for these muscles than had been reported for the cat. Labeled postganglionic sympathetic neurons were found bilaterally in the cranial (superior) cervical, middle cervical and cervicothoracic (stellate) ganglia. Labeled afferent neurons were seen bilaterally in the proximal vagal (jugular) and distal vagal (nodose) ganglia and in the C1-C4 spinal ganglia. The location of sympathetic and sensory nerve cell bodies of the muscles of the laryngopharynx has not been previously reported.

The origins of innervation of the esophagus of the dog.

This study defined the origins of extrinsic efferent and afferent innervation of the normal canine esophagus. When all the layers of the wall of the 3 esophageal regions (cervical, thoracic and abdominal) were injected with horseradish peroxidase (HRP), labeled nerve cells were found in the nucleus ambiguus (NA) and parasympathetic nucleus of X (PX) of the brainstem. Most labeled cells in the NA were located in the compact column (retrofacial nucleus) while labeled cells in the PX were located in separate rostral and caudal areas. There was no somatotopic organization in either the NA or PX. Labeled sympathetic postganglionic neurons were found in the cranial cervical, middle cervical, cervicothoracic, thoracic sympathetic trunk and celiacomesenteric ganglia. The HRP injection of the esophageal wall labeled sensory cell bodies in the glossopharyngeal, proximal and distal vagal, and C2-T6 spinal ganglia. There was no discernible pattern of distribution of labeled cells in the autonomic or sensory ganglia. When the HRP injections were confined to the mucosa-submucosa layers of the thoracic esophagus, a small number of labeled cells were identified in the NA; however, no labeled cells were found in the NA when injections were confined to the mucosa-submucosa of either the cervical or abdominal esophageal regions. With these confined injections, the labeled nerve cells appeared in the rostral part of the PX. Thus, it appeared that the internal tunics of the esophagus (i.e., the mucosa and submucosa) were innervated by neurons in the rostral PX while the muscular tunic was innervated by neurons in the caudal PX and the rostral NA. After mucosa-submucosa injections, labeled sympathetic neurons appeared in the same ganglia that were identified after whole wall injections and these had a similar random distribution. These injections also labeled neurons in the glossopharyngeal, proximal vagal, and distal vagal ganglia, but unlike the whole wall injections there was no labeling in the spinal ganglia. This suggested that the labeled cells of the spinal ganglia seen after whole wall injections conveyed impulses from the tunica muscularis and serosa.

Innervation of the feline eyelids.

The innervation of the eyelids is incompletely understood. This is a particular problem for those who wish to develop animal models of eyelid dysfunction in humans. Blepharospasm, for example, is a disease of uncontrolled eyelid spasm that is difficult to manage clinically because the aetiology is not understood. The anatomical literature on eyelid innervation is sparse and even conflicting. We attempted to study eyelid innervation, both sensory and motor, with injection of horseradish peroxidase (HRP) into the superior eyelid, inferior eyelid, and bulbar conjunctiva. We used 13 anesthetized weanling cats. Shape and structure of the facial nucleus varied along its rostrocaudal extent, but there was a clear demarcation of lateral and medial division. HRP-filled facial nucleus cells were ipsilateral to the injection site, and label appeared throughout the rostrocaudal length. All injection sites, including bulbar conjunctiva, labelled facial nucleus neurons located with overlapping distribution, predominantly in the dorsal part of the lateral division. Likewise, heavy labelling occurred throughout the entire ipsilateral cranial cervical ganglion and the trigeminal ganglion in all kittens. Injection of upper or lower eyelids caused some labelling in the second through the fourth cervical spinal ganglia.

A possible feline model for human blepharospasm.

'Benign essential blepharospasm' is a human eyelid disorder of unknown aetiology characterized by involuntary, bilateral, and disabling spasmodic contracture of the orbicularis oculi muscle. Treatments are frequently disappointing. Here we report what might be a first step toward developing an animal model for exploring mechanisms of the disorder and potential treatments. We surgically implanted stimulating electrodes into brain areas known to supply input to the lateral division of the facial nerve nucleus to induce blinking by electrical stimulation. Single-pulse stimuli at or near the facial, parabrachial, red, and interstitial nuclei produced consistent stimulus-induced eyelid contractions. Responses were ipsilateral to stimulation, except for the interstitial nucleus where contralateral responses occurred. Little or no other movements of the face, head, or body occurred at eye-blink threshold voltages. When these sites were stimulated with pulse trains, eyelid closures followed stimulus frequency and tended to fuse into constant closure. Thresholds at each stimulus site remained constant during three days of testing. Drug treatments produced no consistent effect on eye-blink threshold from any stimulation site, even when general behaviour was affected. We conclude that these input pathways to the facial nucleus may contribute to blepharospasm and that future neurochemical and electrophysiological study of these pathways may produce a suitable animal model for understanding this disorder.

Horseradish peroxidase study of the location of extrinsic efferent and afferent neurons innervating the colon of dogs.

The abdominal portion of the colon of 13 clinically normal dogs was divided into 5 regions (ascending, transverse, left colic flexure, proximal descending, and distal descending), and each region was injected with 30 mg of horseradish peroxidase (HRP). The injected colonic region, brain stem, L7-Cd1 portion of the spinal cord, sympathetic trunk ganglia, celiacomesenteric ganglia, caudal mesenteric ganglion, pelvic plexi, distal vagal (nodose) ganglia, and L1-Cd1 spinal ganglia were obtained at post-injection hour 48, sectioned, and processed by use of the tetramethylbenzidine method. The entire length of the colon was found to be under extrinsic influence of the parasympathetic nucleus of cranial nerve X (PX), with the largest average number of labeled cells resulting from injection of the ascending colon. It was also indicated that the entire colon is under extrinsic influence of the sacral portion of the spinal cord because the pelvic ganglia (second-order neurons) of the pelvic plexi contained labeled cells for all colonic regions. The largest average number of labeled cells in pelvic ganglia was seen after injection of the distal portion of the descending colon. Only after injection of the distal portion of the descending colon were labeled cells found in the S1-S3 portion of the spinal cord. Labeled cells in the PX, spinal cord, and pelvic ganglia were found bilaterally. Although the entire abdominal portion of the colon appears to be influenced by cranial and sacral parasympathetic preganglionic (via pelvic ganglia) neurons, the relative importance of the 2 areas seems to be reversed between the ascending colon and distal portion of the descending colon.(ABSTRACT TRUNCATED AT 250 WORDS)

Shedding of feline immunodeficiency virus in semen of domestic cats during acute infection.

OBJECTIVE: To examine shedding of cell-free and cell-associated feline immunodeficiency virus (FIV) in semen of domestic cats during acute infection. ANIMALS: 7 specific-pathogen-free sexually intact male cats. PROCEDURE: 6 cats were inoculated IV with 5 x 10(6) 50% tissue culture infective doses of FIV-NCSU1, and 1 cat served as an uninfected (control) cat. Infection was confirmed in the 6 cats. Periodically for up to 16 weeks after inoculation, cats were anesthetized and ejaculates obtained by use of electroejaculation. Virus was isolated from filtered seminal plasma and washed seminal cells by co-cultivation with a feline CD4+ T-cell line. Seminal cell lysates were also examined for a 582-base pair segment of FIV gag provirus DNA, using a nested polymerase chain reaction amplification. RESULTS: During the acute phase of FIV infection, virus was evident in semen of 5 inoculated cats. Five cats had virus-positive seminal plasma and 3 had virus-positive cellular constituents during the study. Virus was isolated from 8/22 (36%) seminal plasma samples and 2/17 (18%) seminal cell specimens. Provirus DNA was detected in 5/24 (21%) seminal cell lysates. Cell-free virus was isolated as early as 6 weeks after inoculation, whereas cell-associated virus was isolated as early as 12 weeks after inoculation. Provirus DNA was detected in seminal cells from one cat as early as 1 week after inoculation. CONCLUSIONS AND CLINICAL RELEVANCE: Cell-free and cell-associated FIV are shed in semen of cats early during the course of infection. Samples obtained before seroconversion may contain virus. Virus shedding in ejaculates varies between and within cats during acute infection.

Use of a caudal auricular axial pattern flap in cats.

A caudal auricular axial pattern flap was used in 3 cats to successfully reconstruct large cutaneous defects of the head that were a result of trauma or surgical excision of neoplasms. The technique was a convenient, practical, one-stage procedure for closure of the defects, and flaps up to 7.0 x 12.0 cm were developed. All flaps healed without complications. Anatomic landmarks and surgical guidelines for developing the flap were determined through anatomic dissections and angiographic studies of the cutaneous vasculature of the head and neck in feline cadavers. This report confirms findings of a previous study describing a platysma myocutaneous flap.

Astrocytes and microglia differentially regulate trafficking of lymphocyte subsets across brain endothelial cells.

Feline brain endothelial cells (BECs), astrocytes, and microglia were combined in different configurations in a cell culture insert system to assess the effect of different cell types on the trafficking of peripheral blood mononuclear cell (PBMC) subsets in response to feline immunodeficiency virus (FIV). The addition of astrocytes to BECs significantly increased the adherence of PBMCs. This increase in adherence was suppressed by microglia, whereas microglia alone had no effect on PBMC adherence. FIV exposure of the glial cells did not alter PBMC adherence as compared to same configurations with untreated cells. All PBMC subsets showed some level of trafficking across the endothelial cell layer. The level of trafficking of monocytes and B cells was significantly increased if astrocytes were present. The presence of microglia with the astrocytes reduced transmigration across all PBMC subsets. FIV exposure of astrocytes significantly increased the percentage of CD8 T cell transmigration from 24% to 64% of the total CD4 and CD8 numbers. The presence of microglia significantly reversed the preferential trafficking of CD8 cells in the presence of astrocytes. The results suggested that interaction between the triad of endothelial cells, astrocytes, and microglia played an important, but varying, role in the trafficking of different PBMC subsets. In general, astrocytes had a positive effect on trafficking of PBMCs, while microglia had a suppressive effect. Effects of FIV on trafficking were largely restricted to increases seen in CD8 T cells and monocytes.

Histochemical identification of the striated muscle of the canine esophagus.

Histochemistry on the cervical, thoracic, and abdominal esophageal muscle of immature, young, and adult normal dogs revealed type IIA striated musculature in contrast to some other species. No other types or subtypes were observed. This suggests that esophageal muscle type is established at birth and does not vary or that any variation has been completed by 4 weeks-of-age unlike some canine limb musculature.

The location of extrinsic efferent and afferent nerve cell bodies of the normal canine stomach.

The location of the extrinsic efferent and afferent nerve cell bodies to the mucosa, submucosa, and tunica muscularis of the cardiac, gastric, and pyloric gland regions of the ventral stomach and to the mucosa-submucosa alone of these 3 glandular gastric regions was determined using the horseradish peroxidase technique. All animals of the study demonstrated labeling bilaterally in the rostrocaudal extent of the dorsal motor nucleus of the vagus nerve (DMV) although mucosa-submucosa injections resulted in fewer labeled cells in the DMV. There was no evidence of viscerotopic organization within the DMV for the different gastric regions. However, the left nucleus generally contained a greater number of labeled cells than the right nucleus. Injection of the mucosa, submucosa, and tunica muscularis of the cardiac gland region also resulted in labeling in the nucleus ambiguus in 4 of 5 animals. The vast majority of labeled postganglionic sympathetic neurons were found in the celiacomesenteric ganglion. Labeled cells were also located variously in the stellate ganglion, middle cervical ganglion, and sympathetic trunk ganglia for the different groups. There was no discernible pattern of localization of labeled cells within a sympathetic ganglion. For the stomach, afferent labeled cells were located in the range of the first thoracic to fourth lumbar spinal ganglia and the nodose ganglia, bilaterally. As with sympathetic neurons, there was no discernible pattern of localization of labeled cells within a sensory ganglion.

Percutaneous transtracheal ventilation and morphometric measurements of cervical structures in swine.

Fifteen swine cadavers were examined to determine a quick, reliable means of locating the level of the cricothyroid ligament from consistently palpable structures in the head and neck area. From calculations of measurements made on these cadavers, it was discovered that when a 21-43 kg pig was placed in a standard position, the measured length (L) from the angle of the mandible to the manubrium sterni multiplied by a factor of 0.48 equals the parameter Z [L (0.48) = Z]. Z was measured, beginning at the manubrium sterni, along the ventral midline. The endpoint of Z was at the level of the cricothyroid ligament. A catheter inserted at this point through the ligament into the airway provides a means of transtracheal ventilation. This method of ventilation can be used in emergencies to avoid a tracheostomy or as an alternative to endotracheal ventilation. This method was applied successfully to four anesthetized pigs as determined by a series of arterial blood gas samples.

Persistent oropharyngeal membrane in a Hereford calf.

Persistent oropharyngeal membrane was found in a 6-day-old Hereford calf. The calf was unable to nurse and had not passed feces since birth. Physical examination revealed a fold of tissue spanning the caudal oral cavity, and a barium study demonstrated that food remained within the oral cavity despite a swallowing reflex. A thickened epiglottis was demonstrated radiographically but was not confirmed on necropsy. Necropsy revealed a complete soft-tissue partition between oral cavity and oropharynx with a central blind-ended diverticulum. Only a few cases of persistent oropharyngeal membrane have been reported previously, all in humans. This is the first report of this defect in domestic animals. Possible similarities to the more common persistent anal membrane are discussed.

Symmetric bilateral brachymetacarpalia of a dog.

Malformations of the canine manus are infrequently cited in the literature but include polydactylia and syndactylia. Here, a case of brachymetacarpalia affecting the second and fifth metacarpal bones of both forelimbs of a mixed-breed dog is described. The initial impression of shortened digits was disproved by statistical comparison with normal skeletons, but the presence of shortened metacarpal bones was demonstrated. Other malformations of the manus and appropriate terminology are briefly discussed.

A simplified method of casting the macroscopic airways of lungs.

A technique for preparing casts of the macroscopic airways of mammalian lungs, which is both simplified and inexpensive in comparison with previous techniques, is described. The models are accurate, durable and flexible, and clearly demonstrate the orientation and branching pattern of the bronchial tree. The nature of the procedure also extends the availability of casts to laboratories or individuals with limited instrumentation and/or funding. Preliminary results using this technique to inject the lungs and certain air sacs of birds are also discussed.

Absence of an osseous patella and other observations in Macropodidae stifle.

The patella is a large sesamoid bone that typically develops in the tendon of insertion of the large extensor muscles of the stifle joint. Although present in almost all species of mammals and birds, it has been found to be absent in the red kangaroo and two wallaby species (family Macropodidae). In its place is a fibrocartilage pad, located in the tendon of the quadriceps femoris muscle. This structure is visible grossly, is palpable, and has the form normally expected of a bony patella. In addition, the femoral trochlea is shallow and asymmetric, and the lateral gastrocnemius sesamoid is unusually prominent. These and other related modifications in the area of the Macropodidae stifle are presented.

Cerebrospinal fluid collection in rats: modification of a previous technique.

Modification of a previously published technique for cerebrospinal fluid collection in rats is described. This technique uses general anesthesia, a supporting platform to flex the head-neck junction, a surgical approach to the dorsal atlanto-occipital region, and a micromanipulator to hold and control the approach of the collection needle. Critical steps for success of the procedure are correct positioning, avoiding premature incision of the subarachnoid space, and alignment of the collection needle with the midline and longitudinal axis of the head. The authors had a 95% success rate in obtaining > 0.1 ml of cerebrospinal fluid, using this technique.

Clinical presentation of experimentally induced rabies in horses.

Twelve naive and nine test-vaccinated horses which developed clinical signs of rabies as a result of the required protocol of a vaccine trial were prospectively observed. Nineteen of the 21 cases were confirmed positive for rabies infection of the brain by fluorescent antibody test. The two horses with negative results had ganglioneuritis of the trigeminal ganglion or lymphocytic perivascular cuffing in the brain stem in addition to clinical signs. Average incubation period was 12.3 days and average morbidity was 5.5 days. Naive animals had significantly shorter incubation and morbidity periods (P < 0.05). Muzzle tremors were the most frequently observed (81%) and most common initial sign. Other common signs were pharyngeal spasm or pharyngeal paresis (71%), ataxia or paresis (71%), lethargy or somnolence (71%). The furious form was manifested in 43% of rabid horses and some of these furious animals initially manifested the dumb form. The paralytic form was not observed. Histopathology was characteristics for rabies. The results of this trial do not reflect on the efficacy of commercially licensed equine rabies vaccines.

Clinical features of experimentally induced rabies in cattle and sheep.

A total of 20 cattle and five sheep out of a larger group of animals that were experimentally challenged with virus as part of the required protocol for a vaccine trial developed clinical signs of rabies. All five sheep and 18 of the cattle tested positive for rabies in a direct fluorescent antibody (FA) test. The remaining two cattle had suspicious FA results. Prospective observations are reported in this study. In the diseased cattle, the average incubation period was 15.1 days and the average morbidity period was 3.7 days. Of those, the naive cattle had significantly shorter incubation and morbidity periods than the test-vaccinated cattle. Major clinical signs included excessive salivation (100%), behavioural change (100%), muzzle tremors (80%), vocalization (bellowing; 70%), aggression, hyperaesthesia and/or hyperexcitability (70%), and pharyngeal paresis/paralysis (60%). The furious form of rabies was seen in 70% of the cattle. In the diseased sheep, the average incubation period was 10.0 days and the average morbidity period was 3.25 days. Major clinical signs included muzzle and/or head tremors (80%), aggressiveness, hyperexcitability, and/or hyperaesthesia (80%), trismus (60%), salivation (60%), vocalization (60%) and recumbency (40%). The furious form of rabies manifested in 80% of the sheep. Current rabies vaccines on the market contain higher effective doses than that utilized for the test vaccine and the results of this study do not reflect in any way on commercially available ruminant rabies vaccines.

Choroid plexus macrophages proliferate and release toxic factors in response to feline immunodeficiency virus.

Recent observations have suggested that lentiviruses stimulate the proliferation and activation of microglia. A similar effect within the dense macrophage population of the choroid plexus could have significant implications for trafficking of virus and inflammatory cells into the brain. To explore this possibility, we cultured fetal feline macrophages and examined their response to feline immunodeficiency virus (FIV) or the T-cell-derived protein, recombinant human CD40-ligand trimer (rhuCD40-L). The rhCD40-L was the most potent stimulus for macrophage proliferation, often inducing a dramatic increase in macrophage density. Exposure to FIV resulted in a small increase in the number of macrophages and macrophage nuclei labeled with bromodeoxyuridine. The increase in macrophage density after FIV infection also correlated with an increase in neurotoxic activity of the macrophage-conditioned medium. Starting at 16-18 weeks postinfection, well after the peak of viremia, a similar toxic activity was detected in cerebrospinal fluid (CSF) from FIV-infected cats. Toxicity in the CSF increased over time and was paralleled by strong CD18 staining of macrophages/microglia in the choroid plexus and adjacent parenchyma. These results suggest that lentiviral infection of the choroid plexus can induce a toxic inflammatory response that is fueled by local macrophage proliferation. Together with the observation of increasing toxic activity in the CSF and increased CD18 staining in vivo, these observations suggest that choroid plexus macrophages may contribute to an inflammatory cascade in the brain that progresses independently of systemic and CSF viral load.