Re: [Histonet] lymphatics

From:John Kiernan

Dear Hal Hawkins,

The endothelium of lymphatic vessels can be stained by
enzyme activity histochemistry. The enzyme is a
5-nucleotidase (belongs to the phosphatase family).
Ordinarily this type of method is carried out with frozen
sections of tissue that has been minimally fixed in
formaldehyde or glutaraldehyde, or with unfixed cryostat
sections that have been briefly fixed in cold acetone. The
enzyme in lymphatics may be fairly robust because its
activity can survive embedding in glycol methacrylate. 

The references below may help. If you choose one of these
methods it will be important to consult the original
publication because there's more than one sort of
5-nucleotidase. Each ref is followed by brief notes that I
took when reading the papers. [An R in the acquisition
number means I've got a Reprint of the whole paper; an A
usually means I have a copy of the paper's Abstract. No
letter with the acquisition number usually means I've seen
the whole paper and took notes in the library.] 

8194R. Kato,S; Yasunaga,A; Uchida,U (1991):
Enzyme-histochemical method for identification of lymphatic
capillaries. Lymphology 24, 125-129.
  Glycol methacrylate-embedded sections stained for
5'-nucleotidase & alkaline phosphatase. 5-N-ase in lymph
capills; AlkP-ase in blood capills.

9239R. Nishida,S; Ohkuma,M (1993): Enzyme-histochemical
staining of dermal lymphatic capillaries by guanylate
cyclase. Lymphology 26, 195-199.
  Enzyme histochemical staining distinguishes lymph from
blood capillaries. Says 5-nucleotidase & adenylate cyclase
do so too. (Used unfixed cryostat sections.)

9665R. Okada,E (1994): An improved enzyme-histochemical
method for identification of lymphatic capillaries on
paraffin sections. Lymphology 27, Suppl, 732-735.
  Staining of blood & lymph capillaries. Enzyme
histochemistry for 5-nucleotidase in presence of levamisole
for lymph capills; alkaline phosphatase for blood capills.

11114A. Miura,M; Kato,S; von Ludinghausen,M (1998):
Lymphatic drainage of the cerebrospinal fluid from monkey
spinal meninges with special reference to the distribution
of the epidural lymphatics. Arch. Histol. Cytol. 61(3, Aug),
   5'-nucleotidase staining for lymphatics; alk phosphatase
for blood capillaries (Kato et al '91,'93). Also traced
carbon particles from cisterna magna. Lymphatics and carbon
found on surfaces of cervical & thoracic (most at brachial
plexus levels) roots; not lumbosacral. Epidural lymphatics
most developed on dorsal surface of lower cervical dura.

I hope this helps.
John A. Kiernan
Department of Anatomy and Cell Biology
The University of Western Ontario
London,   Canada   N6A 5C1
"Hawkins, Hal K." wrote:
> For a model of necrotizing enterocolitis in the mouse we need to
> distinguish small veins from lymphatics in the gut.  Could someone
> remind me of any immunostains or lectin that will allow us to tell the
> two apart?
> Many thanks,
> Hal Hawkins, UTMB Galveston
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