Journal of the neurological Sciences, 1975, 25 : 65 74

65

i~ Elsevier Scientific Publishing Company, A m s t e r d a m - Printed in The Netherlands

Protein Patterns of Cerebrospinal Fluid in Hereditary Ataxias and Hereditary Spastic Paraplegia K. G. KJELLIN AND H. STIBLER Department (?['Neurology, Karolinska Hospital, S-104 Ol Stockholm 60 (Sweden) (Received 28 October, 1974)

INTRODUCTION

The hereditary ataxias and allied disorders have been subjected to extensive clinical and pathological studies. In only a few syndromes have biochemical abnormalities been shown. A complex disturbance of lipid metabolism occurs in the BassenKornzweig syndrome (Salt, Wolff, Lloyd, Fosbrook, Cameron and Hubble 1960: Schwartz, Rowland, Eder, Marks, Osserman, Hirschberg and Anderson 1963). In ataxia telangiectasia a deficiency of immunoglobulins, especially IgA has been shown (Gutman and Lemli 1963 ; Fireman, Boesman and Gitlin 1964). In Refsum's disease phytanic acid accumulates in serum and tissues (Wilson and Thompson 1972). In most of these disorders, however, biochemical abnormalities are not yet recognised. The cerebrospinal fluid (CSF) findings have hitherto mostly been reported as normal. In Refsum's disease, however, very high total protein concentrations often occur (Refsum 1946). Lowenthal (1964) and Laterre, Callewaert, Heremans and Sfaello (1970) found normal CSF-protein patterns on agar gel electrophoresis in hereditary ataxias. Normal findings in degenerative diseases were noted also by Stenuit and Delmotte (1972) on paper and agar gel electrophoresis. Laterre and Heremans (1962) noted that in "spinocerebellar and cerebral degenerative diseases", examined by agar electrophoresis, "the post gamma-globulin fraction was conspicuously present" in a few cases. Recently Vandvik and Skrede (1973) reported "mostly normal" findings on agarose gel electrophoresis in 18 unspecified cases with degenerative disorders but they "'occasionally observed prominent bands corresponding to the tau-, beta-trace and gamma-trace localization". Isoelectric focusing is a new technique with high resolving capacity for proteins, which are separated according to their isoelectric points. One procedure for thinlayer isoelectric focusing of CSF-proteins in multiple sclerosis was reported by Delmotte (1971, 1972) and another by Kjellin and Vesterberg (1972). The CSF-protein patterns in some demyelinating and infectious neurological diseases were described by Kjellin (1973) and Kiellin and Vesterberg (1972, 1973, 1974). More recently Kjellin and Stibler (1974a, b) and Stibler and Kjellin (1974) reported the CSF-protein

66

K, G . K J E L L I N ,

[1. S T I B L E R

patterns in extrapyramidal disorders, particularly in Huntington's chorea and spasmodic torticollis. The very high resolving capacity for isoelectric focusing of CSF-proteins, established also from examinations in degenerative neurological diseases as mentioned above, suggested that investigation of the CSF-proteins in hereditary ataxias and hereditary spastic paraplegia would be worthwhile. A preliminary report was given in the abstract by Stibler and Kjellin (1974).

MATERIALS

AND METHODS

The CSF-samples were collected from 12 patients with hereditary ataxias and hereditary spastic paraplegia, treated at the Department of Neurology, Karolinska Hospital, Stockholm. The assessment of the clinical data was based upon examination of the clinical records and the patients were thoroughly examined by one of the authors (H.S.) before the lumbar puncture. The clinical classification was according to Greenfield (1954). The clinical data are given in Table 1. The CSF was obtained by lumbar puncture, performed in a standardized way. About 10 ml of CSF was withdrawn at one lumbar puncture. In addition to isoelectric focusing and quantitative paper electrophoresis the CSF was examined for cells and protein concentration. Serum samples were collected in parallel with the lumbar CSF and examined in a similar way. The cells in the CSF were examined bv routine microscopy and counted per 3.2 mmQ The CSF-protein concentration was determined by a modification of the method of Lowry, Rosebrough, Farr and Randall (1951). The quantitative paper electrophoretic examinations of CSF and serum were performed as described previously {see Ggtrde and Kjellin 1971, including references). The results of the CSF electrophoresis were classified according to characteristic TABLE

1

C L I N I C A L I)ATA O I 12 PATIENTS W I T H Ilt:I~.EI}ITAR~ ArAXIAS ANI) H E R E D I r A R Y SPASFI(

Diagnosis

Friedreich's ataxia

Sex

Patient

LA. ] F.A.

sibs

().A. G.A.J

PARAPLI,(II&

Duration (#' illness (years)

4ge ~years )

Degree qf disabititj

40 50 45

44 53 4o

-t + + + + ÷ + + i.

40

43

+ ~ ~

Holmes' ataxia

O.S.

Marie-Sanger-Brown's ataxia

I.D.

20

25

I.R \ I

25 55

~.2 5,)

5

Hereditary spastic paraplegia

-~

lint,thor of I.R)

I \1. i

sibs

;4?

+ -

ILh. i L.\

17 4

39 3; 2 47

+ {+} 4- ( + }

'-, ~ } ( u n c e r t a i n hcredit), t

5,5

59

i

-

67

PROTEIN PATTERNS OF CSF IN HEREDITARY ATAXIAS

"patterns" of their probable combinations, after correction for the estimated influence of the serum protein distribution (Kjellin 1969) and the CSF-protein patterns were labelled with the symbols given by G~rde and Kjellin (1971). The CSF samples were ultrafiltered (G~rde and Kjellin 1971) and concentrated about 10 times, at which stage samples were withdrawn for isoelectric focusing, and the samples were then further concentrated to about 100 times for quantitative paper electrophoresis. The amounts (20-40 #1) of the concentrated CSF used for isoelectric focusing were selected in order to get similar protein contents in all CSF samples examined. From each patient a serum sample diluted 10 times with physiological NaCI solution was applied side by side with the corresponding concentrated CSF sample on the acrylamide gel used for isoelectric focusing. TABLE 2 THE ISOELECTRIC F O C U S I N G P A T T E R N S ( a - f ) OF CSF G A M M A G L D B U L I N S I ) I V E R G I N G FROM THE " ' N O R M A L " F I N D I N G S IN 2 5 C O N T R O L CASES. V I S U A L E V A L U A T I O N W H E R E THE C O R R E S P O N D I N G S E R U M - P R O T E I N P A T T E R N S HAVE BtiEN IAKI \ I x, I ~ ( O N S I I ) E R A T I O N

Pattern

Relative increase o[ proteins in the re.qions 7-10 (see Fi 9. 1 ) 8-10,especially 9-10 8-10, rather uniformly 7 10, especially 8-10 8 9, rather uniformly 7-9, especially 9 7-9, rather uniformly, usually very similar to a "finger-print" of the corresponding serumgammaglobulin pattern

0

........

U/51

.

Fig. 1. The isoelectric focusing patters (a-f) of CSF gammaglobulins as defined in Table 2. Black and hatched areas indicate relative increase of proteins, where hatched areas indicate a less pronounced increase than in black areas in the patterns a, c and e.

550 480 460 360 440

300 340 550

L.A. F.A. O,A_. G.A. O.S.

I.D. I.R. A.L.

I.M. H.N. L.A. S.O.

Friedreich's ataxia

Holmes" ataxia

Marie Sanger-Brown's ataxia

Hereditary spastic paraplegia

unremarkable unremarkable B, slight unremarkable

unremarkable D. slight-moderate B, slight

unremarkable

B, slight B, slight B, very slight ~ unremarkablc "~

PELF

IF

a a a a

i-.,,c!ccllic Iocu