ABSTRACT
Reissner's fibre (RF) is formed by the polymerization of the glycoprotein secreted by the subcommissural organ (SCO). The SCO also secretes soluble glycoprotein into the cerebrospinal fluid (CSF); variations in RF and SCO have been reported in hydrocephalus. On the other hand, hydrocephalus and other brain alterations have been described in p73 mutant mice. The p73 belongs to the tumour suppressor p53 protein family and has two isoforms: the TAp73 with apoptotic activity and DeltaNp73 with anti-apoptotic function. Moreover, the TAp73 isoform is glycosylated and secreted into the CSF. In the present work, we analysed the variations in RF and p73 proteins in the CSF and SCO of spontaneously hydrocephalic rats. Brains from control rats and spontaneously hydrocephalic rats of 12 months of age were used. The SCO sections were immunohistochemically processed with anti-TAp73 and anti-Reissner fibre (AFRU). The spontaneous hydrocephalus presents a decrease in the AFRU immunoreactive material in the SCO and an absence of RF. The anti-TAp73 was also present, slightly decreased, in the hydrocephalic SCO. AFRU and p73 bands were also detected in the CSF by western blot and six AFRU and p73 protein bands of a similar molecular weight were found in the CSF of the control rats. The number of AFRU and p73 bands was lower in the hydrocephalic rats than in the control rats. In conclusion, hydrocephalus produces a decrease in the secretions of the SCO and an absence of RF and a decrease in p73 and RF proteins in the CSF.
Subject(s)
Cell Adhesion Molecules, Neuronal/cerebrospinal fluid , DNA-Binding Proteins/cerebrospinal fluid , Hydrocephalus/veterinary , Nuclear Proteins/cerebrospinal fluid , Rats, Inbred WKY , Rodent Diseases/cerebrospinal fluid , Subcommissural Organ/metabolism , Tumor Suppressor Proteins/cerebrospinal fluid , Animals , Blotting, Western/veterinary , Case-Control Studies , Cell Adhesion Molecules, Neuronal/analysis , DNA-Binding Proteins/analysis , Hydrocephalus/cerebrospinal fluid , Hydrocephalus/metabolism , Immunohistochemistry/veterinary , Nuclear Proteins/analysis , Rats , Rodent Diseases/metabolism , Subcommissural Organ/chemistry , Tumor Protein p73 , Tumor Suppressor Proteins/analysisABSTRACT
This report describes a new technique for obtaining cerebrospinal fluid from the living mouse (SJL/NBom) in a model of herpes simplex virus encephalitis which is also applicable to other mouse models. The puncture technique was performed in living animals which had been infected with Herpes Simplex Virus Type I strain F in the living animal. The cisterna magna was micro-surgically prepared: The neck muscles were dissected microscopically down to the dura which subsequently was punctured by a glass micropipette. This newly developed minimally invasive technique was performed in a group of living animals (n = 20) and results compared with those of a second group of perfusion fixed animals (n = 20). For the first time, repeated cerebrospinal fluid punctures of individual, living animals are possible. This is of great value for the assessment of new therapeutic and diagnostic strategies in experimental research using mouse models. In addition, this refined methodology significantly reduces the number of experimental animals.
Subject(s)
Cisterna Magna/surgery , Encephalitis, Viral/veterinary , Herpes Simplex/veterinary , Punctures/veterinary , Rodent Diseases/cerebrospinal fluid , Animals , Cerebrospinal Fluid/chemistry , Disease Models, Animal , Encephalitis, Viral/cerebrospinal fluid , Encephalitis, Viral/diagnosis , Female , Herpes Simplex/cerebrospinal fluid , Herpes Simplex/diagnosis , Mice , Microsurgery/methods , Microsurgery/veterinary , Punctures/methods , Rodent Diseases/diagnosis , Specimen Handling/veterinaryABSTRACT
Cerebrospinal fluid (CSF) was removed from guineapigs by puncture of the cisterna magna and the total sample volume of 200-360 microl divided into 40 microl aliquots. After determination of albumin and IgG in these CSF aliquots it was found that successive samples gave different results. In general, up to 100 microl CSF could be removed before the protein concentration began to increase. In animals with chronic relapsing experimental allergic encephalomyelitis (CR-EAE) the rise in albumin concentration was accompanied by a corresponding fall in the number of white cells in later samples.