OBJECTIVETo construct a rat model of chronic nonbacterial prostatitis (CP) and investigate the difference in the quantitative expression of voltage-dependent calcium channels of prostate smooth muscle cells (PSMCs) between the models and controls.

METHODSWe established a CP rat model by estrogen induction, cultured and purified the PSMCs in vitro, and extracted total RNA by Trizol. Then we measured the mRNA expression of the cal subunit in the calcium channel subtypes by reverse transcription and SYBR Green I real time RT-PCR, and compared it with that of the controls.

RESULTSThe expressions of the L-, T- and P/Q-type calcium channels were found in both the CP and control groups, and that of the CaV1.2 L-type calcium channel was significantly increased in the former as compared with the latter (0.048 +/- 0.024 versus 0.031 +/- 0.015, t = 2.846, P = 0.007), but there were no statistically significant differences in the T- and P/Q-type calcium channels between the two groups.

CONCLUSIONThe number of CaV1.2 L-type calcium channels of PSMCs and calcium influx were increased in CP patients, which may be involved in the mechanism of CP.

Animals ; Calcium Channels, L-Type ; metabolism ; Calcium Channels, Q-Type ; metabolism ; Calcium Channels, T-Type ; metabolism ; Estradiol ; pharmacology ; Male ; Myocytes, Smooth Muscle ; metabolism ; Prostate ; metabolism ; Prostatitis ; metabolism ; RNA, Messenger ; genetics ; Rats ; Rats, Wistar

OBJECTIVE: To investigate the effects of CACNA1H gene knockout (KO) on autistic-like behaviors and the morphology of hippocampal neurons in mice. METHODS: In the study, 25 CACNA1H KO mice of 3-4 weeks old and C57BL/6 background were recruited as the experimental group, and 26 wild type (WT) mice of the same age and background were recruited as the control group. Three-chamber test and open field test were used to observe the social interaction, anxiety, and repetitive behaviors in mice. After that, their brain weight and size were measured, and the number of hippocampal neurons were observed by Nissl staining. Furthermore, the CACNA1H heterozygote mice were interbred with Thy1-GFP-O mice to generate CACNA1H^-/--Thy1⁺(KO-GFP) and CACNA1H^+/+-Thy1⁺ (WT-GFP) mice. The density and maturity of dendritic spines of hippocampal neurons were observed. RESULTS: In the sociability test session of the three-chamber test, the KO mice spent more time in the chamber of the stranger mice than in the object one (F_{1, 14}=95.086, P < 0.05; Post-Hoc: P < 0.05), without any significant difference for the explored preference index between the two groups (t=1.044, P>0.05). However, in the social novelty recognition test session, no difference was observed between the time of the KO mice spend in the chamber of new stranger mice and the stranger one (F_{1, 14}=18.062, P < 0.05; Post-Hoc: P>0.05), and the explored preference index of the KO mice was less than that of the control group (t=2.390, P < 0.05). In the open field test, the KO mice spent less time in the center of the open field apparatus than the control group (t=2.503, P < 0.05), but the self-grooming time was significantly increased compared with the control group (t=-2.299, P < 0.05). Morphological results showed that the brain weight/body weight ratio (t=0.356, P>0.05) and brain size (t=-0.660, P>0.05) of the KO mice were not significantly different from those of the control group, but the number of neurons were significantly reduced in hippocampal dentate gyrus compared with the control group (t=2.323, P < 0.05). Moreover, the density of dendritic spine of dentate gyrus neurons in the KO-GFP mice was significantly increased compared with the control group (t=-2.374, P < 0.05), without any significant difference in spine maturity (t=-1.935, P>0.05). CONCLUSION CACNA1H KO mice represent autistic-like behavior, which may be related to the decrease in the number of neurons and the increase in the density of dendritic spine in the dentate gyrus.
Animals ; Autistic Disorder/genetics* ; Calcium Channels, T-Type/genetics* ; Gene Knockout Techniques ; Hippocampus ; Mice ; Mice, Inbred C57BL ; Mice, Knockout ; Neurons

OBJECTIVETo study the relationship of varicocele (VC) with the expressions of T-type channel alpha1H and alpha1G in the sperm of VC patients.

METHODSBased on the WHO criteria, we examined the semen samples by computer-aided sperm analysis (CASA), and divided the samples into groups A (normal semen from volunteers, n = 20), B (normal semen from VC patients, n = 16) and C (abnormal semen from VC patients, n = 44). We optimized the semen by discontinuous Percoll grade centrifugation, and determined the mRNA expressions of T-type channel alpha1H and alpha1G in the three groups using using reverse transcription polymerase chain reaction (RT-PCR).

RESULTSCompared with group A, the mRNA expressions of alpha1H and alpha1G showed with no significant decrease in group B (P>0.05), but were remarkably reduced in group C (P<0.01).

CONCLUSIONThe abnormal mRNA expressions of T-type channel alpha1H and alpha1G may be one of the causes of declined semen quality and consequently infertility in VC patients, which has pointed out a new direction for the studies of the causes and treatment of VC-related infertility.

Adolescent ; Adult ; Calcium Channels, T-Type ; genetics ; metabolism ; Case-Control Studies ; Humans ; Infertility, Male ; genetics ; Male ; RNA, Messenger ; genetics ; Semen Analysis ; Spermatozoa ; metabolism ; Varicocele ; genetics ; metabolism ; Young Adult

OBJECTIVEChildhood absence epilepsy (CAE) is one of the most frequently recognized syndromes among the idiopathic generalized epilepsies (IGEs). It is considered to be a hereditary disease. The possible inheritance pattern of CAE is polygenic. The genes responsible for CAE, however, have not yet been identified. The aim of this study was to further investigate based on the authors' recent work whether or not T-type calcium channel gene-CACNA1H is a susceptibility gene to childhood absence epilepsy.

METHODSThe authors conducted the mutation screening of the exons 6-12 and the nearby partial introns of the CACNA1H gene using the method of direct sequencing of PCR products in 48 newly found CAE patients.

RESULTSThe authors found 13 single nucleotide polymorphisms (SNPs). They also found 4 mutations which only existed in CAE patients. Both G773D and H515Y mutations were heterozygous. The mutation of H515Y has never been reported previously. The patient inherited the mutation from his mother. The authors found two CAE patients with the mutation of G773D previously. This is the third time that the authors found one more CAE family with this G773D mutation, and the patient with the mutation G773D inherited the mutation from his father.

CONCLUSIONT-type calcium channel gene-CACNA1H might be a susceptibility gene to childhood absence epilepsy.

Amino Acid Sequence ; Calcium Channels, T-Type ; genetics ; Child ; Child, Preschool ; Epilepsy, Absence ; genetics ; Genetic Predisposition to Disease ; Humans ; Molecular Sequence Data ; Mutation ; Polymorphism, Single Nucleotide

OBJECTIVETo observe the anti-remodeling effect of acupuncture on asthma and to explore the mechanism of T-type calcium channel protein in airway smooth muscle cell in airway remodeling effect in asthma.

METHODSThirty-two rats were randomly divided into a normal group, a model group, an acupuncture group and a sham acupuncture group, 8 rats in each group. The rats in the latter three groups were sensitized for consecutive 14 days by single peritoneal injection of aqueous suspension 1 mL of 10 mg ovalbumin (OVA), 200 mg aluminum hydroxide and saline together with 1 mL inactivated pertussis vaccine. From the 15th day, asthma was induced for 30 minutes by ultrasonic atomizing inhalation of 1% OVA for consecutive 14 days in the model group. The acupuncture group was treated with acupuncture at "Dazhui" (GV 14), "Fengmen" (BL 12) and "Feishu" (BL 13) for 30 minutes before the ultrasonic atomizing inhalation, once every two days for consecutive 14 days. The same acupoints selection and the course of treatment as the acupuncture group were produced in the sham acupuncture group and they were treated with acupuncture at 1 mm acupoint skin without retaining needles. The normal group remained unhandled. The respiratory function and the airway remodeling were evaluated by airway resistance and pulmonary histopathology, respectively, and the T-type calcium channel protein expression of Ca(v)3.1, Ca(v) 3.2, Ca, 3.3 in airway smooth muscle cell were detected by immunohistochemistry technique.

RESULTS(1) The airway resistance in the model group was higher than that in the normal group and in the acupuncture group (both P < 0.05), and the airway resistance in the acupuncture group was lower than that in the sham acupuncture group (P < 0.05). (2) The ratios of the airway wall thickness to the basement membrane perimeter (Awt/Pbm) and the airway outer perimenter to the airway internal perimeter (Po/Pi) in the model group were higher than those in the normal group and in the acupuncture group (all P < 0.05), and the ratios of Awt/Pbm and Po/Pi in the acupuncture group were lower than those in the sham acupuncture group (both P < 0.05). (3) The average optical of Ca(v) 3.1 and Ca(v) 3.2 in airway smooth muscle cell in the model group were higher than that in the normal group and in the acupuncture group (both P < 0.05), and the average optical of Ca(v) 3.3 in airway smooth muscle cell in the model group was higher than that in the normal group (P < 0.05) and it was lower than that in the sham acupuncture group (P < 0.05).

CONCLUSIONAcupuncture can inhibit the airway remodeling and the accrementition of the airway smooth muscle and can reduce the airway resistance. The mechanism may be related to the inhibition of T-type calcium channel protein in airway smooth muscle cell, especially in relation to the protein expression of Ca(v) 3.1.

Acupuncture Therapy ; Airway Remodeling ; Animals ; Asthma ; genetics ; metabolism ; physiopathology ; Calcium Channels, T-Type ; genetics ; metabolism ; Humans ; Male ; Myocytes, Smooth Muscle ; metabolism ; Rats ; Rats, Sprague-Dawley ; Respiratory System ; metabolism ; physiopathology

OBJECTIVETo study the effect of CACNA1H gene mutation G773D on calcium channel function.

METHODSBy the overlap extension PCR we introduced G773D mutation into a human Cav3.2acDNA for constructing the mutant. And then using whole cell clamp technique, we studied the alterations of channel behavior in transfected HEK-293 cells.

RESULTSThere were no difference in kinetics of activation and inactivation of calcium channel between wild type and mutant. However comparing with the wild-type Cav3.2 channel, G773D mutant could increase the calcium current density significantly.

CONCLUSIONCACNA1H gene G773D mutation is able to increase calcium current and neuronal excitability.

Base Sequence ; Calcium Channels, T-Type ; genetics ; physiology ; Cell Line ; Child ; Child, Preschool ; DNA Mutational Analysis ; Epilepsy, Absence ; genetics ; pathology ; physiopathology ; Family Health ; Female ; Humans ; Male ; Molecular Sequence Data ; Mutation ; Patch-Clamp Techniques ; Polymorphism, Single Nucleotide