Streptozotocin-induced liver damage in mice

Diabetes incidence and liver damage was studied and identified in C3H-s mice 21 days after Streptozotocin (SZ) administration (250 mg/kg/i.v.) at 04 hs (4 a.m.) and 16 hs (4 p.m.). Metabolic disturbances were assessed by daily control of glycosuria and serum glucose determined at the end of the experiment. Liver damage was controlled by light and electron microscopy. Both effects showed a circadian variation, with significant greatest values in the 16-h-injected group. Liver damage appeared whether the animals became diabetic or not, consisting in degranulation of the rough-surfaced endoplasmic reticulum, mitochondrial swelling with loss of cristae and edema of the ground substance, with flocculent amorphous precipitate. In some hepatocytes, a dilated cisternae of the endoplasmic reticulum was seen. It was concluded that: a) beta-cell and hepatocytes have a synchronic circadian sensitivity to SZ; b) liver damage was present whether the animals became diabetic or not, suggesting the presence of a different threshold for SZ effect in hepatocytes. These results might be taken into account when planning SZ use, either for experimental or clinical purposes.     

Teratogenic effect of ketamine and cocaine in CF-1 mice

Pregnant CF-1 mice were used to study the teratogenic effect of ketamine and cocaine, alone and in combination. The dose of ketamine was 50 mg/kg and that of cocaine was 20 mg/kg, given intravenously (tail) once daily (these doses of ketamine and cocaine are comparable to doses used by addicted humans). Treatment was started from day 6 to day 15 of gestation, and dams were sacrificed on day 18. There were significant decreases in the fetal weight and length in the combined group. Skeletal defects such as incomplete ossification of skull bones and vertebrae were observed in both the cocaine and combined group, compared with the control. An increased frequency of cerebral and abdominal hemorrhages as well as hydrocephalus and hydronephrosis was observed in the combined group. This study showed that fetal exposure to ketamine and cocaine in combination was more teratogenic than each drug alone in CF-1 mice.     

Teratogenic effect of cocaine and diazepam in CF1 mice

This study was conducted to determine the teratogenic effect of cocaine hydrochloride, alone or in combination with diazepam. Pregnant mice were administered cocaine hydrochloride at 10 and 20 mg/kg body weight intravenously (tail), and/or diazepam at 20 or 40 mg/kg b.w. by gavage. Combinations of diazepam and cocaine (20/10 and 40/20, respectively) were used. Significant reductions of fetal weight and length were found in the group treated with diazepam and cocaine (40/20). The incidences of incomplete ossification of sternebrae and skull bones, as well as delayed ossification of the paws, were significantly increased in the combination groups. Hydronephrosis and cryptorchidism were observed in the cocaine-treated groups, with the incidence of these malformations increasing significantly when diazepam was co-administered.     

Bromobenzene causes Clara cell damage in mice

The oral administration of a single dose of bromobenzene (5 mmol/kg) causes cellular damage in the lungs of C57B1/6 male mice. At 24 h following treatment with bromobenzene, Clara cells of the bronchiolar epithelium demonstrated necrosis; endoplasmic reticular cisternae were extensively dilated, and degenerated membranes were present in the cytoplasmic matrix as electron-dense tubular aggregates. The ciliated cells which were interspersed among the Clara cells, remained unaffected. The morphologic alterations produced by bromobenzene and the target cell are similar in character to the pulmonary lesions produced by administration of other aromatic and aliphatic halogenated hydrocarbons.     

Proliferative response and renewal of hepatic function following cocaine administration in mice

Experiments were undertaken to investigate the hepatic, temporal and spatial sequence of events following a single injection of cocaine, a known hepatotoxin. Centrilobular necrosis was induced in male mice (DBA/2Ha) 24 hr post-injection (PI). The time course of hepatic damage was monitored by assaying microsomal cytochrome P450 content, the activity of microsomal FAD-containing monooxygenase (FAD-M) and by determining the levels of serum glutamic pyruvic transaminase (SGPT). Kinetics of the onset of DNA synthesis were determined by autoradiography of thin liver sections and the incorporation of 3H-methyl thymidine into perchloric-acid-precipitable material. There was no increase in the labelling index (LI) and thymidine (TdR) incorporation in the first 24 hr PI. The LI rose to 14.6% and TdR incorporation showed a 5-fold increase over control values 48 hr PI. Both indices declined slightly at 72 hr PI and returned to control values by 96 hr PI. In contrast, the cytochrome P450 content declined by 69%, the FAD-M activity dropped by 40% and the SGPT levels showed an 18-fold increase at 24 hr PI, coincident with cytological signs of necrosis. Although the patterns of recovery differed between these selected enzymes, normal values were attained by 96 hr PI. These results demonstrate that cell damage and hepatic dysfunction precede the onset of DNA synthesis and subsequent proliferation.     

Mitochondrial dysfunction and oxidative damage in parkin-deficient mice

Loss-of-function mutations in parkin are the predominant cause of familial Parkinson's disease. We previously reported that parkin-/- mice exhibit nigrostriatal deficits in the absence of nigral degeneration. Parkin has been shown to function as an E3 ubiquitin ligase. Loss of parkin function, therefore, has been hypothesized to cause nigral degeneration via an aberrant accumulation of its substrates. Here we employed a proteomic approach to determine whether loss of parkin function results in alterations in abundance and/or modification of proteins in the ventral midbrain of parkin-/- mice. Two-dimensional gel electrophoresis followed by mass spectrometry revealed decreased abundance of a number of proteins involved in mitochondrial function or oxidative stress. Consistent with reductions in several subunits of complexes I and IV, functional assays showed reductions in respiratory capacity of striatal mitochondria isolated from parkin-/- mice. Electron microscopic analysis revealed no gross morphological abnormalities in striatal mitochondria of parkin-/- mice. In addition, parkin-/- mice showed a delayed rate of weight gain, suggesting broader metabolic abnormalities. Accompanying these deficits in mitochondrial function, parkin-/- mice also exhibited decreased levels of proteins involved in protection from oxidative stress. Consistent with these findings, parkin-/- mice showed decreased serum antioxidant capacity and increased protein and lipid peroxidation. The combination of proteomic, genetic, and physiological analyses reveal an essential role for parkin in the regulation of mitochondrial function and provide the first direct evidence of mitochondrial dysfunction and oxidative damage in the absence of nigral degeneration in a genetic mouse model of Parkinson's disease.     

Proteasome inhibition attenuates coxsackievirus-induced myocardial damage in mice

Coxsackievirus B3 (CVB3) is one of the most prevalent pathogens of viral myocarditis, which may persist chronically and progress to dilated cardiomyopathy. We previously demonstrated a critical role of the ubiquitin-proteasome system (UPS) in the regulation of coxsackievirus replication in mouse cardiomyocytes. In the present study, we extend our interest to an in vivo animal model to examine the regulation and role of the UPS in CVB3-induced murine myocarditis. Male myocarditis-susceptible A/J mice at age 4-5 wk were randomized to four groups: sham infection + vehicle (n = 10), sham infection + proteasome inhibitor (n = 10), virus + vehicle (n = 20), and virus + proteasome inhibitor (n = 20). Proteasome inhibitor was administered subcutaneously once a day for 3 days. Mice were killed on day 9 after infection, and infected hearts were harvested for Western blot analysis, plaque assay, immunostaining, and histological examination. We showed that CVB3 infection led to an accumulation of ubiquitin conjugates at 9 days after infection. Protein levels of ubiquitin-activating enzyme E1A/E1B, ubiquitin-conjugating enzyme UBCH7, as well as deubiquitinating enzyme UCHL1 were markedly increased in CVB3-infected mice compared with sham infection. However, there was no significant alteration in proteasome activities at 9 days after infection. Immunohistochemical staining revealed that increased expression of E1A/E1B was mainly localized to virus-damaged cells. Finally, we showed that application of a proteasome inhibitor significantly reduced CVB3-induced myocardial damage. This observation reveals a novel mechanism of coxsackieviral pathogenesis, and suggests that the UPS may be an attractive therapeutic target against coxsackievirus-induced myocarditis.     

Conditioned place preference for cocaine and methylphenidate in female mice from lines selectively bred for high voluntary wheel-running behavior

Behavioral addictions can come in many forms, including overeating, gambling and overexercising. All addictions share a common mechanism involving activation of the natural reward circuit and reinforcement learning, but the extent to which motivation for natural and drug rewards share similar neurogenetic mechanisms remains unknown. A unique mouse genetic model in which four replicate lines of female mice were selectively bred (>76 generations) for high voluntary wheel running (High Runner or HR lines) alongside four non-selected control (C) lines were used to test the hypothesis that high motivation for exercise is associated with greater reward for cocaine (20 mg/kg) and methylphenidate (10 mg/kg) using the conditioned place preference (CPP) test. HR mice run ~three times as many revolutions/day as C mice, but the extent to which they have increased motivation for other rewards is unknown. Both HR and C mice displayed significant CPP for cocaine and methylphenidate, but with no statistical difference between linetypes for either drug. Taken together, results suggest that selective breeding for increased voluntary running has modified the reward circuit in the brain in a way that increases motivation for running without affecting cocaine or methylphenidate reward.