Calretinin Immunoreactivity Research Articles

Distribution of the Cocaine and Amphetamine - Regulated Transcript (Cart) and Calcium Binding Proteins Immunoreactivity in the Preoptic Area of the Ram

Abstract The aim of the study was to describe the distribution of cocaine and amphetamine-regulated transcript (CART) and calcium binding proteins (CaBPs) of EF-hand family, namely calbindin, calretinin, and parvalbumin in the preoptic area (POA) of the ram. Frozen sections were processed for a routine immunofluorescence labelling. CART, calbindin, and calretinin immunoreactivity was present in neurons and fibers of the preoptic area, whereas parvalbumin showed immunoreactivity only in the POA fibers. CART displayed from a moderate to low immunoreactivity in cells and a high immunoreactivity in fibers. The highest immunoreactivity of all studied CaBPs exhibited calbindin, whereas the lowest parvalbumin. The results of the present study suggest that among the studied CaBPs, calbindin is the most likely to be involved in the participation of the important regulatory functions in the ram’s POA and the rich CART innervation seems to be strictly related to its control of the reproduction.

Calretinin Immunohistochemistery: An Aid in the Diagnosis of Hirschsprung's Disease.

Definite diagnosis of Hirschsprung's disease (HD) is based on histopathological study, but there are limitations associated with standard histology and histochemistry in this regard. The aim of this study was to investigate calretinin immunostaining patterns in both ganglionic and aganglionic HD intestinal specimens and to compare them with control specimens. Specimens included 30 patients with histopathologic diagnosis of HD and 20 patients that underwent colectomy for other reasons (as control group). Eighty paraffin wax blocks of full thickness intestinal specimens (30 blocks of ganglionic segments, 30 blocks of aganglionic segments and 20 blocks of control group) were studied. Calretinin immunoreactivity and pattern of staining for ganglion cells (nuclear and cytoplasmic) and also nerve fibers in different layers of bowel were evaluated in IHC stained slides. There were positive immunostaining of nerve fibers in the lamina propria, submucosa and muscularis propria in control and patient group. There were also nuclear and cytoplasmic staining of ganglion cells in submucosa and muscularis propria in all specimens of both control group (100%) and ganglionic segments (100%). Calretinin immunoexpression of nerve fibers in muscularis propria of the aganglionic segments was negative in all but two cases (6.7%). This method had sensitivity of 93.3% and specificity of 100% for diagnosis of HD in full thickness specimens of intestinal wall. The positive predictive value was 100% and negative predictive value was 93.8%. Calretinin immunohistochemistry can be used on suction rectal biopsies as a reliable and adjunctive method to diagnose HD.

Immunohistochemical Study of Calretinin in Normal Skin and Cutaneous Adnexal Proliferations

Calretinin is a calcium-binding protein member of the EF-hand family. The presence of calretinin has been demonstrated in certain stages of the cellular cycle in a wide variety of normal and neoplastic tissues. The main aims of our study were (1) to investigate what structures of the normal skin and cutaneous adnexal proliferations express immunoreactivity for calretinin and (2) to determine the value of immunohistochemical expression for calretinin as a marker for follicular, sebaceous, apocrine, and eccrine differentiation in cutaneous adnexal proliferations. We studied 139 biopsy specimens, including 10 cases of normal skin of different locations and 129 benign and malignant cutaneous adnexal proliferations. In normal skin, we found that calretinin is expressed in the innermost cell layer of the outer root sheath in anagen hair follicle, in both the duct and sebolemma of the sebaceous gland, in the secretory portion of eccrine glands, and in mast cells of the stroma. In cutaneous adnexal proliferations, we found strong immunoreactivity for calretinin in tricholemmal cysts, tricholemmomas/inverted follicular keratoses, tumors of follicular infundibulum, and in some basal cell carcinomas. Focal positivity was also seen in trichoadenomas, trichoblastomas/trichoepitheliomas, pilomatricomas, proliferating tricholemmal tumors, pilar sheath acanthomas, trichofolliculomas, follicular hybrid cysts, cutaneous mixed tumors, steatocystomas, sebaceous hyperplasias, and sebaceomas. These results demonstrate that immunohistochemical study for calretinin may be helpful to identify the innermost cell layer of the outer root sheath in anagen hair follicle and the cutaneous adnexal proliferations showing differentiation toward this structure. Calretinin immunoreactivity supports eccrine differentiation in some sweat gland neoplasms, and it is also useful in identifying neoplasms with ductal sebaceous differentiation.

Mesotheliomas with small cell features: report of eight cases

AbstractMesotheliomas with small cell morphology are rare and only one study of such cases has been published. As a result of their rare occurrence, some investigators have cast doubt on the existence of such a histologic variant of mesothelioma. This investigator reports a series of eight cases of epithelioid mesothelioma with small cell features, all of which originated in the pleura. Seven of the patients were men and one was a woman. Four patients had a history of asbestos exposure. Histologically, four of the mesotheliomas were epithelioid and four biphasic. The proportion of small cells seen in these cases constituted 80 to 100% of the tumor included in the biopsy material and 15 to 20% of the tumor present in the pneumonectomy specimens. Immunoreactivity for calretinin, keratin 5/6, keratin 7, pan-keratin, WT1, podoplanin, and mesothelin was seen in all cases tested for these markers. All of the cases were negative for MOC-31, Ber-EP4, CEA, CD15, TAG-72, TTF-1, chromogranin A, synaptophysin, CD99, and desmin. The mean survival of the six patients for whom this information was available was 8.2 months. It is important for pathologists to be aware that mesotheliomas can present small cell features and, because of this, they can be confused with other malignancies that can exhibit similar morphology. The value of immunohistochemistry in the differential diagnosis of these tumors is discussed.

Age-related changes in calbindin and calretinin immunoreactivity in the central auditory system of the rat

Age-related changes in the levels of major intracellular calcium buffers are known to occur in different parts of the mammalian brain, including the central auditory pathway. In the present study, we evaluate with immunohistochemistry and the western blot technique the effect that aging has on the calbindin- and calretinin-expressing system of neurons in the higher structures of the central auditory pathway, in the inferior colliculus (IC), medial geniculate body (MGB) and auditory cortex (AC) of two rat strains, the slowly aging Long-Evans and the fast aging Fischer 344. Interestingly, the age-related changes demonstrated a similar character regardless of the rat strain. In the IC of young animals, the majority of calbindin and calretinin immuno-reactive (CB and CR-ir) cells were found in the dorsal and external cortices and only sparse positive cells were present in the central nucleus of the IC. With aging, the number of CB-ir and CR-ir neurons decreased significantly in both the dorsal and external cortices. Furthermore, these declines were accompanied by an age-related reduction in the mean volumes of CB- and CR-ir neuronal somas. In the MGB of young rats, CB-ir neurons were present in abundant numbers in both the dorsal and ventral subdivisions, while CR-ir neurons were practically absent in this structure. With aging, the number and mean volume of CB-ir cells in the ventral subdivision of the MGB were significantly decreased. In comparison with the IC and MGB, age-related numerical and volumetric declines of both CB-ir and CR-ir neurons in the AC were less pronounced. Western blot protein analysis revealed a pronounced age-related decline in the levels of calbindin in both strains and in all examined brain regions. In contrast, the decline in calretinin levels with aging was less prominent, with a significant decline only in the IC of both strains. The observed age-related changes in the calbindin- and calretinin-expressing systems may contribute significantly to the deterioration of hearing function known as central presbycusis.

The changes of calretinin immunoreactivity in paraquat-induced nephrotoxic rats

Calcium-binding proteins are present in the kidneys: calbindin D-28k in the distal tubules and calretinin in the proximal tubules. Since paraquat causes degeneration in the brush border-bearing proximal tubule cells in rat kidneys, we investigated the changes of calretinin immunoreactivity in the proximal tubule cells of paraquat-induced nephrotoxicity in experimental male Sprague-Dawley rats following chitosan oligosaccharide pretreatment to investigate its protective properties. Paraquat (60mg/kg) was administered intraperitoneally with or without chitosan oligosaccharide (500mg/kg, p.o.) pretreatment. The changes on calretinin were compared with those of calbindin D-28k by immunohistochemistry and Western Blot analysis. Calretinin was immunolocalized on the apical surface of proximal tubule cells in the deeper cortex of normal kidney, and disappeared after paraquat administration with minor changes of calbindin D-28k immunoreactivity in the distal tubules and collecting ducts. Chitosan oligosaccharide pretreatment caused increased expression of calretinin and calbindin D-28k before paraquat injection and helped preserve proximal tubules after paraquat treatment. However, Western blot analysis on calretinin and calbindin D-28k could not explain the degeneration of the proximal tubule cells in paraquat-induced nephrotoxicity. These findings suggested that calretinin is a possible and more useful histopathological marker for proximal tubule cells in paraquat-induced nephrotoxic rats.

Immunocytochemical Analysis of Calretinin in the Frontal Cortex of Chinchilla

Abstract The aim of the study was to define morphology and distribution of calretinin (CR) positive neurons in the frontal cortex of adult chinchilla males and intracellular localisation of the protein in this area. The brains of 5 adult chinchilla males were used in the study. CR immunoreactive neurons were shown with peroxidase-antiperoxidase immunohistochemical reaction using a specific monoclonal antibody. Intensive CR immunoreactivity was demonstrated mainly in few polymorphic neurons of II, III, and V layers. Cytoplasmic and nuclear reaction product in most CR positive neurons was diffuse and in some neurons of layer V in the form of granules localised peripherally. In a few cells more intensive staining was observed in the nuclei than in the cytoplasm. The results indicate the presence of heteromorphic CR positive neurons in specific layers of the frontal cortex. Nuclear localisation of CR in neurons suggests passive transport of this protein, which may affect the nuclear genes. This protein is a neuroprotector maintaining appropriate level of calcium, modulating neuronal activity, and synaptic conduction in the frontal cortex of chinchilla.

Pericardial Synovial Sarcoma, a Potential for Misdiagnosis

Synovial sarcomas arising in unexpected locations may lead to diagnostic challenges. In this report, we describe 3 cases of synovial sarcoma that manifested clinically as primary pericardial lesions. All 3 cases occurred in men in their fourth decade. Fever, cough, chest pain, and chest distress were the most common symptoms. Histologically, 2 of the tumors were spindle cell monophasic, and 1 tumor was biphasic. By immunohistochemical studies, the tumor cells were positive for cytokeratins and epithelial membrane antigen. In addition, the tumor cells displayed focal immunoreactivity for calretinin, cytokeratin 5/6, and HBME-1, resulting in the initial interpretations of malignant mesotheliomas. None of the 3 cases were diagnosed correctly until subsequent molecular cytogenetic assays demonstrated the presence of SYT gene rearrangements. As there are overlapping morphologic features between pericardial synovial sarcoma and mesothelioma, molecular analysis is essential for differential diagnoses.

Calretinin, parvalbumin and calbindin immunoreactive interneurons in perirhinal cortex and temporal area Te3V of the rat brain: Qualitative and quantitative analyses

The perirhinal cortex (PRC) composed of areas 35 and 36 forms an important route for activity transfer between the hippocampus–entorhinal cortex and neocortex. Its function in memory formation and consolidation as well as in the initiation and spreading of epileptic activity was already partially elucidated. We studied the general pattern of calretinin (CR), parvalbumin (PV) and calbindin (CB) immunoreactivity and its corrected relative optical density (cROD) as well as morphological features and density of CR and PV immunoreactive (CR+, PV+) interneurons in the rat PRC. Neighboring neocortical association area Te3V was analyzed as well.The PRC differed from the Te3V in higher CR and lower PV overall immunoreactivity level. On CR immunostained sections, the difference between high cROD value in area 35 and low cROD value in area Te3V reached statistical significance (p<0.05). The pattern of CB immunoreactivity was similar to that of the neocortex. Vertically oriented bipolar neurons were the most common morphological type of CR+ neurons, multipolar neuronal morphology was typical among PV+ neurons and vertically oriented bipolar neurons and multipolar neurons were approximately equally frequent among CB+ neurons. The density of CR+ and PV+ neurons was stereologically measured. While the density of PV+ neurons was not significantly different in PRC when compared to Te3V, density of CR+ neurons in area 35 was significantly higher by comparison with Te3V (p<0.05). Further, the overall neuronal density was measured on Nissl stained sections and the proportion of CR+ and PV+ interneurons was expressed as a percentage of the total neurons counts. The percentage of CR+ interneurons was higher in area 35 by comparison with area Te3 (p<0.05), while the percentage of PV+ interneurons did not significantly differ among the examined areas. In conclusion, the PRC possesses specific interneuronal equipment with unusually high proportion of CR+ interneurons, what might be of importance for the presumed gating function of PRC in normal and diseased states.

Calcium-binding proteins and GFAP immunoreactivity alterations in murine hippocampus after 1 month of exposure to 835 MHz radiofrequency at SAR values of 1.6 and 4.0 W/kg

Widespread use of wireless mobile communication has raised concerns of adverse effect to the brain owing to the proximity during use due to the electromagnetic field emitted by mobile phones. Changes in calcium ion concentrations via binding proteins can disturb calcium homeostasis; however, the correlation between calcium-binding protein (CaBP) immunoreactivity (IR) and glial cells has not been determined with different SAR values. Different SAR values [1.6 (E1.6 group) and 4.0 (E4 group) W/kg] were applied to determine the distribution of calbindin D28-k (CB), calretinin (CR), and glial fibrillary acidic protein (GFAP) IR in murine hippocampus. Compared with sham control group, decreased CB and CR IRs, loss of CB and CR immunoreactive cells and increased GFAP IR exhibiting hypertrophic cytoplasmic processes were noted in both experimental groups. E4 group showed a prominent decrement in CB and CR IR than the E1.6 group due to down-regulation of CaBP proteins and neuronal loss. GFAP IR was more prominent in the E4 group than the E1.6 group. Decrement in the CaBPs can affect the calcium-buffering capacity leading to cell death, while increased GFAP IR and changes in astrocyte morphology, may mediate brain injury due to radiofrequency exposure.

Hypothyroid States Mitigate the Diabetes-Induced Reduction of Calbindin D-28k, Calretinin, and Parvalbumin Immunoreactivity in Type 2 Diabetic Rats

In this study, we investigated the differences in calbindin D-28k (CB), calretinin, (CR) and parvalbumin (PV) immunoreactivity in the hippocampus of Zucker diabetic fatty (ZDF) rats and Zucker lean control (ZLC) rats. In addition, we observed the effects of hypothyroidism on the levels of immunoreactivity of these proteins in ZDF rats. For this study, 7-week-old ZDF rats were used, and methimazole treatment was continued for 5 weeks to induce hypothyroidism. The animals were sacrificed at 12 weeks of age. ZDF rats showed increased blood glucose levels compared to those in ZLC rats. Methimazole intervention significantly reduced total and free T3 levels, and it ameliorated the increase of blood glucose levels in ZDF rats. In ZLC rats, CB, CR, and PV immunoreactivity was detected in regions of the hippocampus proper. In vehicle-treated ZDF rats, CB, CR, and PV immunoreactivity was significantly decreased in the hippocampus. However, in the methimazole-treated rats, CB, CR, and PV immunoreactivity was significantly increased compared to that in the vehicle-treated rats. These results suggest that hypothyroidism ameliorated the diabetes-induced reduction of CB, CR, and PV immunoreactivity in the hippocampus.

Medioventral part of the posterior thalamus in the mouse

The posterior thalamus (Po) consists of heterogeneous groups of cells, which have not been clearly defined. In the present study, we focused on a part of the Po in the mouse brain, which is located caudally to the ventral posterior nucleus and rostromedially to the medial geniculate nucleus and shows distinct calretinin immunoreactivity. While we found the region had a considerable unity on the cytoarchitectural and histochemical grounds, it did not correspond to any particular nucleus but partially involved three structures in a widely used brain atlas ( Franklin and Paxinos, 2008). Therefore, we tentatively designated the region as the medioventral part of the posterior thalamus (PoMV) and examined its anatomical features with immunohistochemistry and retrograde tract-tracing. The PoMV was appreciated as a reticular structure with prominent calretinin immunoreactivity, especially in horizontal sections, and displayed apparent differences in the cytoarchitecture from its surrounding regions. The PoMV had two divisions: the dorsal division (PoMVd), which contained parvalbuminimmunoreactive fibers, and the ventral division (PoMVv), which lacked these fibers. The tract-tracing studies showed that the somata retrogradely labeled from the injections in the insular cortex and some of the extended amygdalar regions were fairly concentrated within the PoMV, especially in the PoMVd. On the other hand, the labeling from the medial hypothalamus injections was found predominantly within the PoMVv. These findings indicate that the PoMV can be regarded as a distinct structure within the Po, and it may play a role in the emotional aspect of somatosensory processing.

Effect of Ginseng on Calretinin Expression in Mouse Hippocampus Following Exposure to 835 MHz Radiofrequency

Exponential rise in the use of mobile communication devices has generated health concerns due to radiofrequency (RF) exposure due to its close proximity to the head. Calcium binding proteins like calretinin regulate the levels of calcium (Ca2+) which plays an important role in biological systems. Ginseng is known for maintaining equilibrium in the human body and may play a beneficial radioprotectant role against electromagnetic field (EMF) exposure. In the present study, we evaluated the radioprotective effects of red ginseng (RG) extract in a mouse model. Calretinin (CR) expression was measured using a free-floating immunohistochemical method in the hippocampus of mice after 835 MHz EMF exposure for 5 h/d for 5 d at specific absorption rate=1.6 W/kg for the different experimental groups. The control animals were treated with NaCl while the experimental animals received 10 mg/kg ginseng, or 30 mg/kg; EMF exposed mice were also treated with NaCl, 10 mg/kg ginseng (E10), or 30 mg/kg (E30). Decreases in CR immunoreactivity (IR) along with loss of CA1 and CA3 interneurons and infragranular cells were observed in the ENaCl group while such losses were not observed in the E10 and E30 groups. CR IR significantly increased in the RG-treated group compared to control and EMF-exposed groups treated with NaCl. The study demonstrates that RG extract can serve as a radioprotective agent that maintains Ca2+ homeostasis and prevents neuronal loss in the brain hippocampal region caused by RF exposure.

Effects of Ischemia and Reperfusion on P2X2 Receptor Expressing Neurons of the Rat Ileum Enteric Nervous System

We investigated the effects of ischemia/reperfusion in the intestine (I/R-i) on purine receptor P2X2-immunoreactive (IR) neurons of the rat ileum. The superior mesenteric artery was occluded for 45 min with an atraumatic vascular clamp and animals were sacrificed 4 h later. Neurons of the myenteric and submucosal plexuses were evaluated for immunoreactivity against the P2X2 receptor, nitric oxide synthase (NOS), choline acetyl transferase (ChAT), calbindin, and calretinin. Following I/R-i, we observed a decrease in P2X2 receptor immunoreactivity in the cytoplasm and surface membranes of neurons of the myenteric and submucosal plexuses. These studies also revealed an absence of calbindin-positive neurons in the I/R-i group. In addition, the colocalization of the P2X2 receptor with NOS, ChAT, and calretinin immunoreactivity in the myenteric plexus was decreased following I/R-i. Likewise, the colocalization between P2X2 and calretinin in neurons of the submucosal plexus was also reduced. In the I/R-i group, there was a 55.8% decrease in the density of neurons immunoreactive (IR) for the P2X2 receptor, a 26.4% reduction in NOS-IR neuron, a 25% reduction in ChAT-IR neuron, and a 47% reduction in calretinin-IR neuron. The density of P2X2 receptor and calretinin-IR neurons also decreased in the submucosal plexus of the I/R-i group. In the myenteric plexus, P2X2-IR, NOS-IR, ChAT-IR and calretinin-IR neurons were reduced in size by 50%, 49.7%, 42%, and 33%, respectively, in the I/R-i group; in the submucosal plexus, P2X2-IR and calretinin-IR neurons were reduced in size by 56% and 72.6%, respectively. These data demonstrate that ischemia/reperfusion of the intestine affects the expression of the P2X2 receptor in neurons of the myenteric and submucosal plexus, as well as density and size of neurons in this population. Our findings indicate that I/R-i induces changes in P2X2-IR enteric neurons that could result in alterations in intestinal motility.

Differential Effects of Treadmill Exercise on Calretinin Immunoreactivity in Type 2 Diabetic Rats in Early and Chronic Diabetic Stages

In this study, we investigated the effects of treadmill exercise on calretinin (CR), a marker of early postmitotic neurons, immunoreactivity in the dentate gyrus (DG) of Zucker diabetic fatty (ZDF) rats, before or after diabetes onset, and Zucker lean control (ZLC) rats. For this study, 6-week-old ZLC and prediabetic ZDF rats, and 22-week-old ZLC and ZDF rats were exercised on the treadmill. Sedentary ZLC and ZDF rats of the same age were used as exercise experiment controls. The exercised prediabetic ZDF rats did not show diabetes onset, while the sedentary prediabetic ZDF rats showed significantly increased blood glucose levels. The exercised diabetic ZDF rats exhibited a decrease in their blood glucose levels compared to the sedentary diabetic ZDF rats, but the levels were still above 20 mmol/l. ZLC rats of both ages were in the normoglycemic range. CR immunoreactivity was detected throughout the DG, including the subgranular zone and the polymorphic layer. Diabetic rats exhibited a significant decrease in the number of CR-immunoreactive cells and fibers in the DG. Exercise in the prediabetic ZDF rats significantly increased the number of CR-immunoreactive cells and fibers in the subgranular zone of the DG. In the ZLC and ZDF rats of chronic diabetic phase, exercise increased CR-immunoreactive neurons in the hilar region. These results suggest that diabetes significantly reduces the number of postmitotic CR-immunoreactive neurons and the intensity of immunoreactivity and that exercise increases these CR-related parameters in a diabetic stage-dependent manner.

Peritoneal malignant psammomatous mesothelioma

Psammoma bodies (PBs) are observed most commonly in papillary thyroid carcinoma, meningioma, and papillary serous cystadenocarcinoma of the ovary. We report one case of peritoneal malignant mesothelioma (PMM) with massive deposition of PBs. A 72-years-old man presented with abdominal swelling and marked weight loss. Contrast enhanced computed tomography showed fluid diffuse in peritoneal recesses, thick septa with micronodules in the greater omentum and adjacent enhancement of the thickened peritoneum. The explorative laparoscopy showed diffuse minute parietal peritoneal nodules. The peritoneal biopsy revealed a superficial papillary growth of malignant epithelial-like cells with diffuse involvement of submesothelial tissues. Massive deposition of PBs was observed. Nuclear and cytoplasmic calretinin immunoreactivity was present in neoplastic cells along with membranous D2-40 and membranous/cytoplasmic cytokeratin 5 staining. The patient was treated with chemotherapy (gemcitabine, vinorelbine, cisplatin). PBs may represent an active biologic process ultimately leading to degeneration/death of tumor cells and retardation of growth of the neoplasm. It may also serve as a barrier against the spread of tumor. Psammomatous malignant mesothelioma may simulate serous psammocarcinoma of the peritoneum. The behavior of serous psammocarcinoma is more closely similar to borderline serous tumor than to serous carcinoma. Further studies are necessary to establish if massive deposition of PBs may define a new variant of psammomatous malignant mesothelioma with a favorable impact to the prognosis of usual psammomatous malignant mesothelioma, as well as in serous psammocarcinoma of the peritoneum.

Calretinin immunoreactivity in normal and carbon tetrachloride-induced nephrotoxic rats

Carbon tetrachloride (CCl 4) is a potent hepatotoxic and nephrotoxic chemical. Little, however, is known about the association of CCl 4-induced nephrotoxicity and calretinin. We hypothesized that calretinin might be localized in the proximal tubule cells and play a role against CCl 4-induced nephrotoxicity, since the target of CCl 4 is the brush border-bearing tubule cells. CCl 4 (1 ml/kg) was administrated by oral gavage to 8-week old male Sprague–Dawley rats once a week for 4 weeks. A significant increase in serum blood urea nitrogen and creatinine was confirmed by serum analysis. Calretinin immunolocalization was compared with the calbindin D-28k immunoreactivity in normal and CCl 4-treated kidneys. Calretinin was clearly immunolocalized in the apical surface of proximal convoluted tubule in the deeper cortex of normal kidney and blurred after CCl 4 administration, with only minor changes of calbindin D-28k immunoreactivity in the distal convoluted tubules and collecting ducts, irrelevant to the CCl 4 treatment. These findings might have significance since decreased immunolocalization of calretinin with CCl 4-induced nephrotoxicity may contribute to the toxicity-related decrease in calcium transport or calcium buffering activity in the kidney.

Long‐term regulation in calretinin staining in the rat inferior colliculus after unilateral auditory cortical ablation

In this study we analyzed the effects in the inferior colliculus of a unilateral ablation of the auditory cortex in rats. Variations in both calretinin immunoreactivity and protein levels determined by Western blot suggest that such lesions induce changes in the regulation of this calcium-binding protein. Stereological counts of calretinin-immunoreactive neurons in the inferior colliculus 15, 90, and 180 days after the lesion showed a progressive increase in the number of immunoreactive neurons, with a parallel increase in the intensity of staining. Two hundred forty days after the cortical lesion, both the number of immunoreactive neurons and the staining intensity had returned to control values. The effects of the cortical lesion on calretinin regulation are more intense in those inferior colliculus subdivisions more densely innervated by the corticocollicular projection. This finding, along with the time course of calretinin regulation suggests that degeneration of the descending projection is linked to calretinin regulation in the inferior colliculus. We hypothesize, based on the role of calretinin, that the observed increase in immunoreactivity levels seen in the inferior colliculus after lesioning of the auditory cortex may be related to altered excitability in deafferented neurons. Our finding, may reflect adaptive mechanisms to changes in calcium influx and excitability in inferior colliculus neurons induced by lesions of the descending projection from the cortex to the inferior colliculus.

Immunohistochemical localization of calbindin D28-k, parvalbumin, and calretinin in the cerebellar cortex of the circling mouse

The spontaneous mutant circling mouse has an autosomal recessive pattern of inheritance and is an animal model for deafness, which is characterized by circling, head tossing, and hyperactivity. Since the main pathology in circling mice lies in the organ of Corti, most studies on deaf mice have focused on auditory brain stem nuclei. No studies regarding behavior-related CNS changes in circling mice have been reported. The major center of sensory input for modulation of motor activity is best-studied in the cerebellum. Considering the importance of calcium homeostasis in numerous processes, calcium-binding proteins (CaBPs), such as calbindin D-28k (CB), parvalbumin (PV), and calretinin (CR), may play crucial roles in preserving cerebellar coordinated motor function. Thus, the distribution of CB, PV, and CR was determined in the cerebellum using immunohistochemical methods to compare immunoreactivity (IR) of CaBPs between wild-type (+/+), heterozygous (+/cir), and homozygous (cir/cir) mice. The IR of CB and PV was predominantly observed in the Purkinje cell layer of all three genotypes. Compared with the +/+ genotype, the relative mean density of CB and PV IR in the Purkinje cell layer and CR IR in the granular layer was significantly decreased in the cir/cir genotype. Changes in calcium homeostasis in parallel fiber/Purkinje cell synapses could diminish cerebellar control of motor coordination. A number of deficiencies among the CaBPs lead to distinct alterations in brain physiology, which may affect normal behavior.

Sclerochorioretinal Abnormalities in Hypercholesterolemic Rabbits Treated with Rosiglitazone

To evaluate early retinal, choroidal, and scleral abnormalities induced by a hypercholesterolemic diet and the prevention of these abnormalities after oral administration of rosiglitazone in rabbits. Fifty-four New Zealand rabbits were divided into four study groups: control group, normal diet; group 1, hypercholesterolemic diet; group 2, hypercholesterolemic diet associated with daily administration of 3 mg of rosiglitazone from day 14 after beginning the diet; and group 3, hypercholesterolemic diet associated with daily administration of 3 mg of rosiglitazone since the beginning of the experiment. Sclera and choroid underwent histologic and histomorphometric analyses. Retina underwent immunohistochemical analysis with anti-calretinin and anti-glial fibrillary acidic protein (GFAP) antibodies. No abnormalities were observed in the control group. Group 1 had significant increases in scleral and choroidal thicknesses compared with the control group (P < .01) and group 3 (P < .05). Group 1 presented significant increases in immunoreactivity (P < .001) to the anti-calretinin antibody compared with the other groups. Groups 2 and 3 had significant (P < .002) increases in calretinin immunoreactivity compared with the control group. GFAP was negative in all groups. The hypercholesterolemic diet induced early retinal, choroidal, and scleral abnormalities. Rosiglitazone preserved the structural anatomy.