BACKGROUNDWe investigated the co-expression of calbindin-D28k (CB), calretinin (CR) and parvalbumin (PV, a combination of the three is referred to as CaBPs) with gamma-aminobutyric acid (GABA) or glycine in neurons of the rat medullary dorsal horn (MDH).

METHODSImmunofluorescence histochemical double-staining for CaBPs and GABA or glycine was performed on the sections from rat MDH.

RESULTSCB-, CR-, PV-, GABA- and glycine-like immunoreactive (LI) neurons were differentially observed in all layers of the MDH, but particularly in lamina II. Neurons that exhibited immunoreactivity for both CaBPs and GABA or glycine were also observed mainly in lamina II. A few of them were found in laminae I and III. The percentages of neurons which co-expressed CB/GABA or CB/glycine out of the total numbers of CB- and GABA-LI neurons or CB- and glycine-LI neurons were 5.3% and 12.1% or 4.1% and 10.0%, respectively. The ratios of CR/GABA or CR/glycine co-existing neurons out of the total numbers of CR- and GABA-LI neurons or CR- and glycine-LI neurons were 5.8% and 7.6% or 4.4% and 7.1%, respectively. The rates of PV/GABA or PV/glycine co-localized neurons out of the total numbers of PV- and GABA-LI neurons or PV- and glycine-LI neurons were 11.1% and 5.1% or 9.9% and 5.1%, respectively.

CONCLUSIONThe results indicate that some neurons in the MDH contain both CaBPs and GABA or glycine.

Animals ; Calbindin 1 ; Calbindin 2 ; Calbindins ; Calcium-Binding Proteins ; analysis ; Fluorescent Antibody Technique ; Glycine ; analysis ; Immunohistochemistry ; Medulla Oblongata ; cytology ; Parvalbumins ; analysis ; Posterior Horn Cells ; chemistry ; Rats ; S100 Calcium Binding Protein G ; analysis ; gamma-Aminobutyric Acid ; analysis

The maturation pattern of the calcium binding proteins parvalbumin (PV) and calbindin-D28K (CB) from the day of birth, postnatal day 0 (P0) to 30 days (P5, P10, P15, P20, P30) and adult was studied in the rat amygdala using immunohistochemistry. PV and CB immunoreactivities in the amygdala of the rat showed very different patterns during postnatal development. The first PV-immunoreactive neurons appeared in the cortical amygdaloid nuclei and the basolateral amygdaloid nucleus at P5, and then in the lateral amygdaloid nucleus and the basomedial amygdaloid nucleus at P10. Adult patterns of PV-immunoreactive neurons were reached at P20. In contrast, CB-immunoreactive neurons were already found at birth in all amygdaloid nuclei except the intercalated nucleus. The intensity and number of immunoreactivity of CB-containing neurons increased during the first 10 days of postnatal life but dramatically decreased at P15. Mature patterns CB-immunoreactive neurons were achieved at P20. These two calcium binding proteins exhibited a non-homogeneous distribution in the adult amygdala, PV-immunoreactive neurons were mainly localized in the basolateral nuclear group but not in the medial amygdaloid nucleus, the cental amygdaloid nucleus and the intercalated nucleus. In contrast, CB-immunoreactive neurons were distributed in almost all amygdaloid nuclei except the intercalated nucleus. The present results showing different postnatal maturation patterns such as time of appearance, the number and distribution of immunoreactive cells suggest that PV and CB may play a different functional role during the postnatal development of the amygdala.
Adult ; Amygdala ; Animals ; Calbindin 1 ; Calbindins* ; Calcium-Binding Proteins ; Humans ; Immunohistochemistry ; Neurons* ; Parturition ; Rats*

The interstitial nucleus of the spinal trigeminal tract (INV) contains many calbindin-D28k-containing neurons (CB-neurons) receiving convergence information from the somatic and visceral structures. The purpose of the present study was to confirm whether the primary afferent terminals from the inferior alveolar nerve (IAN) make close contact and synaptic connections with the same CB-neurons receiving visceral nociceptive signals in INV. Biotinylated dextran amine (BDA) and horseradish peroxidase (HRP) tracing combined with CB and Fos proteins immunohistochemistry were used. After injections of BDA and formalin into unilateral IAN and upper alimentary tract, respectively, the transganglionic labeled afferent fibers and terminals from IAN were observed in the ipsilateral INV, especially in its enlarged part. A large number of CB- and Fos-like immunoreactive (LI) neurons were found in bilateral INV. These CB- and Fos-LI neurons mostly overlapped with BDA-labeled terminals in the enlarged part of INV. About one half of the CB-LI neurons were double labeled with Fos-LI nuclei (74/153). The terminals from IAN were to made close contacts with many CB/Fos-double labeled or CB-single labeled neurons. After injection of HRP into IAN, HRP-labeled fibers and terminals in INV were similar to that labeled with BDA. Under the electron microscope, a large number of CB-LI dendrites and a few soma in the enlarged part of INV were found to form asymmetrical axo-dendritic and axo-somal synapses with the HRP-labeled axon terminals. These results indicate that the orofacial somatic inputs from IAN and the visceral nociceptive inputs from the upper alimentary tract converge onto the same CB-containing neurons in INV. These CB-containing neurons in INV probably play an important role in information integration as well as visceral and cardiovascular activity.
Animals ; Calbindin 1 ; Calbindins ; Face ; innervation ; Male ; Microscopy, Confocal ; Neural Pathways ; cytology ; physiology ; Neurons ; physiology ; Nociceptors ; physiology ; Presynaptic Terminals ; physiology ; Proto-Oncogene Proteins c-fos ; physiology ; Rats ; Rats, Sprague-Dawley ; S100 Calcium Binding Protein G ; metabolism ; physiology ; Trigeminal Nuclei ; physiology ; Viscera ; innervation

It has been reported that new apical anion exchanger perndrin, encoded by the pendred syndrome (PDS/pds, Slc26A4) gene, was expressed in the AE1-negative intercalated cells of rat and mouse kidneys. The purpose of this study was performed that expression of pendrin in the subtypes of intercalated cells in human kidney. The normal human renal tissues obtained from nephrotomized kidneys for renal cell carcinoma were fixed in periodate-lysine-paraformalde-hyde, and processed for immunohistochemistry. Subtypes of intercalated cells were identified by using antibodies for H(+)-ATPase and AE1, and connecting tubule cells and principal cells of collecting duct were identified using antibodies for calbindin D28K and AQP2, respectively. In human kidney, pendrin was expressed in the apical domain of AE1-negative intercalated cells including type B cells with diffuse and/or basolateal H(+)-ATPase, non A-non B (non -A/B) type intercalated cells with apical H(+)-ATPase and bipolar type of intercalated cells with apical and basolateral H(+)-ATPase. The AQP2-positive principal cells of cortical collecting duct were also had apical pendrin immunoreactivity. However, there was no pendrin immunoreactivity in AE1-positive type A intercalated cells, calbindin D28K-positive connecting tubule cells, and AQP2-positive medullary collecting duct. These results suggest that pendrin is an apical anion exchanger not only in the AE1-negative intercalated cells (type B, non-A/B and bipolar cells) but also in the principal cells of cortical collecting duct, and has an essential role in HCO3-secretion in human kidney.
Animals ; Antibodies ; B-Lymphocytes ; Calbindin 1 ; Calbindins ; Carcinoma, Renal Cell ; Humans* ; Immunohistochemistry ; Kidney* ; Mice ; Proton-Translocating ATPases ; Rats

The α2A adrenoceptors (α2A-ARs) are the most common adrenergic receptor subtype found in the prefrontal cortex (PFC). It is generally accepted that stimulation of postsynaptic α2A-ARs on pyramidal neurons are key to PFC functions, such as working memory. However, the expression of α2A-ARs in interneurons is largely unknown. In the present study using double-labeling immunofluorencence technique, we investigated the expression of α2A-ARs in major types of rat PFC interneurons expressing calcium-binding proteins parvalbumin (PV), calretinin (CR), and calbindin (CB). Our data demonstrated that α2A-ARs are highly expressed in calcium-binding protein immunoreactive interneurons of rat PFC, suggesting that stimulation of α2A-ARs may alter neural networks comprising pyramidal neurons and interneurons, thereby exerting a beneficial effect on PFC cognitive functions. The present study provides the morphological basis for a potential mechanism by which stimulation of α2A-ARs induces cognitive improvement.
Animals ; Calbindin 2 ; metabolism ; Calbindins ; metabolism ; Interneurons ; metabolism ; Parvalbumins ; metabolism ; Prefrontal Cortex ; cytology ; Rats ; Receptors, Adrenergic, alpha-2 ; metabolism

Adult ; Biomarkers, Tumor ; metabolism ; Calbindin 2 ; Female ; Follow-Up Studies ; Humans ; Mesothelioma ; metabolism ; pathology ; surgery ; Mucin-1 ; metabolism ; Peritoneal Neoplasms ; metabolism ; pathology ; surgery ; S100 Calcium Binding Protein G ; metabolism

Many researches have focused upon temporal changes of neurotransmitters and/or neuromodulators in the central nervous system after ischemic insult. In sensory neurons, the spatial and temporal alterations of neurotransmitters have been little studied. Calbindin D-28k (CB) and calretinin (CR) have been suggested to play a role in the transmission of neurotransmitters. Therefore, in the present study we investigated the chronological alteration of CB and CR immunoreactivity in the trigeminal ganglion cells of the Mongolian gerbil after ischemic insult. In the sham operated group, CB and CR immunoreactivities were found in small -, medium -and large -sized neurons. One and two days after ischemia-reperfusion, small and large-sized CB immunoreactive neurons increased significantly. Thereafter, number of the CB immunoreactive neurons decreased markedly. Furthermore, five days after ischemia -reperfusion, CB immunoreactivity was detected in a few neurons, and its immunoreactivity was also very weak in the cytoplasm. Number of the large -sized CR immunoreactive neurons increased significantly one day after ischemia -reperfusion. Thereafter, the number of the large -sized CR immunoreactive neurons decreased. Especially, the number of the medium-sized CR immunoreactive neurons increased dramatically 4 days after ischemia-reperfusion. These results suggest that an increase of CB and CR may play an important role in modulating the mechanoception 1 day after ischemia-reperfusion, because the immunoreactivities increased in large -sized neurons which have the myenlinated A fibers. These results also suggest that significant increase of CR expression in medium -sized neurons 4 and 5 days after ischemia-reperfusion may provoke CR in modulating the nociception or thermoception because the medium-sized neurons which have the myenlinated A sigma or C fibers.
Calbindin 2* ; Calbindins* ; Central Nervous System ; Cytoplasm ; Gerbillinae* ; Immunohistochemistry ; Ischemia ; Nerve Fibers, Myelinated ; Nerve Fibers, Unmyelinated ; Neurons ; Neurotransmitter Agents ; Nociception ; Sensory Receptor Cells ; Trigeminal Ganglion*

The synaptic contacts of cochlear afferent fibers (CAFs) with inner hair cells (IHCs) are spatially segregated according to their firing properties. CAFs also exhibit spatially segregated vulnerabilities to noise. The CAF fibers contacting the modiolar side of IHCs tend to be more vulnerable. Noise vulnerability is thought to be due to the absence of neuroprotective mechanisms in the modiolar side contacting CAFs. In this study, we investigated whether the expression of neuroprotective Ca²⁺-buffering proteins is spatially segregated in CAFs. The expression patterns of calretinin, parvalbumin, and calbindin were examined in rat CAFs using immunolabeling. Calretinin-rich fibers, which made up ~50% of the neurofilament (NF)-positive fibers, took the pillar side course and contacted all IHC sides. NF-positive and calretinin-poor fibers took the modiolar side pathway and contacted the modiolar side of IHCs. Both fiber categories juxtaposed the C-terminal binding protein 2 (CtBP2) puncta and were contacted by synaptophysin puncta. These results indicated that the calretinin-poor fibers, like the calretinin-rich ones, were afferent fibers and probably formed functional efferent synapses. However, the other Ca²⁺-buffering proteins did not exhibit CAF subgroup specificity. Most CAFs near IHCs were parvalbumin-positive. Only the pillar-side half of parvalbumin-positive fibers coexpressed calretinin. Calbindin was not detected in any nerve fibers near IHCs. Taken together, of the Ca²⁺-buffering proteins examined, only calretinin exhibited spatial segregation at IHC-CAF synapses. The absence of calretinin in modiolar-side CAFs might be related to the noise vulnerability of the fibers.
Animals ; Calbindin 2* ; Calbindins ; Carrier Proteins ; Fires ; Hair Cells, Auditory, Inner ; Intermediate Filaments ; Nerve Fibers ; Noise* ; Rats ; Sensitivity and Specificity ; Synapses ; Synaptophysin

BACKGROUND AND OBJECTIVES: Food restriction retards aging and increases mean and maximum life span in nearly all species tested thus far. Calcium-binding proteins show a heterogenous distribution in the mammalian central nervous system and are useful markers for identifying neuronal populations. These proteins have been implicated in the buffering and transport of calcium as well as in the regulation of various enzyme systems. We investigated the change of the immunoreactivity of calcium-binding proteins in olfactory bulb of rat after food restriction. MATERIALS AND METHOD: 10 weeks old Sprague-Dawley rat were used in this study. 6 rats were killed at the beginning of the experiment. 30 rats which were restricted food only half of their normal voluntary mean food intake (12 g instead of 24 g per day) were killed at 3 days, 1, 2, 4 and 9 weeks after food restriction (n=6 per time point). Olfactory bulbs of the rats were cut into 40 micro m-thick coronal sections and immunostained. RESULTS: On the layers of olfactory nerve, glomerular, outer plexiform, granular cell and subependymal zone, immunoreactivities of parvalbumin and calbindin were increased on food restriction week 1 and 2. However, parvalbumin at olfactory nerve layer and calbindin at granule cell layer failed to increase at week 2. Calretinin increased its immunoreactivity at olfactory nerve and outer plexiform layer at week 1. After restriction week 2, immunoreactivity of calcium-binding proteins was almost same as control. CONCLUSION: The results we obtained from restricted rats indicated that parvalbumin, calbindin and calretinin could be expressed by different manner and layer in olfactory bulb.
Aging ; Animals ; Calbindin 2 ; Calbindins ; Calcium* ; Calcium-Binding Proteins* ; Central Nervous System ; Eating ; Neurons ; Olfactory Bulb* ; Olfactory Nerve ; Rats* ; Rats, Sprague-Dawley

OBJECTIVE: To analyze the survival and the changes of proportions of Calbindin, Calretinin and Parvalbumin positive neurons in mouse hippocampal CA area at chronic stage of Pilocarpine-induced epilepsy. METHODS: Calbindin, Calretinin and Parvalbumin immunofluoresence staining were done 2 months after Pilocarpine-induced epilepsy in mice or saline injection. RESULTS: Two months after Pilocarine-induced epilepsy, the number of Calbindin, Calretinin and Parvalbumin positive neurons in the CA area decreased significantly compared with the control (P<0.01), especially the Calbindin positive neurons had a great drop and Pavalbumin positive neurons had a least drop. At the chronic stage of epilepsy, the proportion of Calbindin, Calretinin and Parvalbumin positive neurons in the CA area was changed. The content of Pavalbumin positive neurons increased whereas the content of Calbindin positive neurons decreased significantly compared with the control (P<0.01). CONCLUSION The changes of proportions of Calbindin, Calretinin and Parvalbumin positive neurons in the CA area of mouse hippocampus may be a factor in the ongoing epileptic activity at chronic stage of Pilocarpine-induced epilepsy.
Animals ; Calbindin 2 ; metabolism ; Calbindins ; metabolism ; Cell Survival ; physiology ; Chronic Disease ; Epilepsy ; chemically induced ; metabolism ; Hippocampus ; metabolism ; Male ; Mice ; Neurons ; metabolism ; Parvalbumins ; metabolism ; Pilocarpine ; gamma-Aminobutyric Acid ; metabolism