Proteins of rat serum, urine, and cerebrospinal fluid: VI. Further protein identifications and interstrain comparison.

We have investigated the biological fluids--serum, cerebrospinal fluid, and urine--of three strains of rats; the present data extend our database (also available on-line) and may be of interest for pharmacological and toxicological investigation. Specifically, we have defined reference maps of the major protein components in cerebrospinal fluid and urine. Compartment-specific isoforms were recognized for transferrin and transthyretin. Mass spectrometric data established the cleavage site of the signal peptide and identified the N-terminal blocking group of prostaglandin D synthase from rat cerebrospinal fluid. A previously undescribed member of the family of low molecular mass rat urinary proteins was characterized as containing a sequence similar, but not identical, to the N-terminal region of rat urinary protein-2 (RUP-2), and divergent from RUP-1.

Toxicity studies and distribution dynamics of retroviral vectors following intrathecal administration of retroviral vector-producer cells.

The use of intrathecal, retroviral-mediated transfer of the herpes simplex thymidine kinase (HStk) gene and subsequent ganciclovir (GCV) administration has recently been shown to improve survival in a rat model of leptomeningeal carcinomatosis. Clinical application of this approach is attractive because access to the cerebrospinal fluid (CSF) space is relatively noninvasive and distribution of producer cells and vectors may be facilitated by circulation of CSF, overcoming distribution problems inherent in solid tumors. However, meningeal inflammation, transduction and injury to normal CNS tissue, proliferation of the xenogeneic producer cells in the subarachnoid space, immune-mediated injury, and development of hydrocephalus are possible complications of intraventricular or intrathecal administration of vector-producer cells. In addition, the dynamics of producer cell and vector distribution in the CSF are unknown. To address these issues, we evaluated the safety of this approach for gene delivery and assessed the dynamics of distribution of producer cells and retroviral vectors in rats and non-human primates. In rats, transduction of normal central nervous system (CNS) structures surrounding the subarachnoid space was evaluated after intrathecal and intraventricular injections of beta-galactosidase and HStk vector-producer cells, with and without GCV. In primates, beta-galactosidase and HStk vector-producer cells were injected intraventricularly and GCV was administered either intrathecally or intravenously. Toxicity was evaluated by neurologic examination, serial gadolinium-enhanced MRI scans of the brain, and blood and CSF profiles. A subgroup of monkeys received repeated intraventricular injection of vector-producer cells and intravenous GCV. The titer of retroviral-vector was measured in cisternal and lumbar CSF samples after repeated producer cell injection.(ABSTRACT TRUNCATED AT 250 WORDS)

Changes in physiological parameters of rat cerebrospinal fluid during chronic sampling: evaluation of two sampling methods.

Although rat cerebrospinal fluid (CSF) is increasingly being used in pharmacological and biochemical research, methodological studies on basic physiological data are lacking. We have determined the albumin content and number of erythrocytes and leukocytes in CSF obtained by two different methods of sampling from cisterna magna-repeated sampling from an implanted cannula and by repeated punctures. In the initial samples the albumin content was 0.08 +/- 0.03 micrograms/microliters. Chronic cannulation of the cisterna magna resulted in a meningeal reaction with increased cell and albumin content: a reaction that could be reduced but not prevented by using a sterile cannula. The number of leukocytes but not erythrocytes was highly correlated to the albumin content. Repeated sampling in the absence of a permanent cannula did not significantly increase albumin content but carried a higher risk for erythrocyte contamination.

[A new technique for collection the cerebrospinal fluid from rat (authors transl)].

An attempt was made to collect the cerebrospinal fluid (CSF) of rat using a capillary phenomenon technique. Experiments were carried out in 76 rats anesthetized with pentobarbital sodium (Nembutal, Abbot. 50 mg/kg IP). The midline incision in the occipital region was made from 0.5 cm anterior to interauricular line of 4 cm in length. Exposing the atlanto-occipital dura mater by separating the nuchal muscles, the CSF was collected with a heparinized microtube (MICROPET. Bection, Dickson and Company) through a small hole made by a 18G syringe needle. Thus, 0.05-0.05 ml of the CSF could be readily obtained without mixture of the blood in 64 cases out of 76 trials, 84%. This simple technique is a useful way to get a small amount of CSF from little animals, such as rats.