This file includes introduction to multiple sclerosis (MS) , their sign and symptoms , types of multiple sclerosis, pathophysiology of MS , again this includes the medication that are used to treat MS , & the last point is the Preclinical Screening models or methods for multiple sclerosis . Preclinical Screening models includes in vivo and in vitro models

1. PRECLINICAL SCREENING
METHODS FOR
NEURODEGENERATIVE DISEASES (
MULTIPLE SCLEROSIS)
PRESENTED BY:- Janhavi Yashwant
Burade
M.pharm 1st sem
VBCOP , AMRAVATI

2. CONTENT
 INTRODUCTION
 PATHOPHYSIOLOGY
 MEDICATIONS
 SCREENING METHODS

3. INTRODUCTION
Multiple sclerosis is one of the neuro degenerative
disorder in central nervous system that include brain,
spinal cord and nerves.
Including problem with vision ,arm or leg movement,
sensation or balance.

4. SYMPTOMS OF MS
 The symptoms of MS vary widely from person to person and can affect
any part of the body.The main symptoms include:
• fatigue
• difficulty walking
• vision problems, such as blurred vision
• problems controlling the bladder
• numbness or tingling in different parts of the body
• muscle stiffness and spasms
• problems with balance and co-ordination
• problems with thinking, learning and planning

5. TYPES OF MULTIPLE SCLEROSIS (MS)
1. Relapsing-remitting
2. 2. Primary progressive
3. Progressive relapsing
4. Secondary progressive relapsing

6. PATHOPHYSIOLOGY OF MS
 MS is an autoimmune condition, which means your immune
system mistakes part of your body for a foreign substance
and attacks it.
 The immune system attacks axon, leads to destruction of
myelin sheath in brain and Spinal cord resulting in
Conduction block which leads to permanent loss of function.
 The attacks cause the myelin sheath to become inflamed in
small patches (plaques or lesions),which can be seen on an
MRI scan
 These patches of inflammation can disrupt the messages
travelling along the nerves.

8. MEDICATION
 Azathioprine
 Cyclophosphamide
 Dalfampridine
 Dexamethasone
 Dimethyl fumarate
 Fingolimod
 Glatiramer
 Mitoxantrone
 Natalizumab
 Prednisolone
 teriflunomide

9. SCREENING MODELS
IN-VIVO
•Experimental autoimmune
encephalomyeltis
•Viral
•Toxin models
•Transgenic, mutant and parabiotic
mice
IN-VITRO
•Microglia
•Oligodendrocytes
•Astrocytes
•Neurons
•Brain Slice and Aggregate Systems
•Blood-Brain Barrier Models

10. IN-VITRO
Microglia
1. Microglia activation is observed in actively demyelinating MS lesions,
pre-active lesions, areas of remyelination as well as the normal-
appearing white matter.
2. primary microglia cultures are derived from embryonic or early post-
natal animals.
PROCEDURE
 This method is simple and allows for relatively high yields of cells.
 Microglia can also be separated from confluent primary mixed glial
cultures by agitation on a rotary shaker, producing a highly enriched (>
95%) cell culture.
 Given that microglia in the brain , The initial trigger of microglia activation.
 microglia phenotype supportive of regeneration is observed at the
earliest stages of demyelination

11. IN-VIVO MODELS
1.EXPERIMENTAL AUTO IMMUNE ENCEPHALOMYLTIS
1. EAE is a spectrum of neurological disorders induced in
laboratory animals following immunisation with CNS.
2. Antigens emulsified in an adjuvant to augment the
immune response
3. These models generally use purified myelin, recombinant
proteins or encephalitogenic peptides of myelinproteins.
4. EAE studies used myelin basic protein (MBP) since it is a
major protein component of the myelin sheath and highly
soluble, and can therefore be purified relatively easily.

12. PROCEDURE :-
 Active EAE relies on CNS-reactive T cells that are induced
following immunisation by an autoantigen
 Emulsified in an adjuvant, while passive or adoptive transfer
EAE is induced by transferring the autoreactive T cells to
native recipient animals.

13.  Adjuvants used for EAE induction frequently contain killed
mycobacteria that elicit T cell responses as well as
antibody production due to innate immune activation
viaToll-like receptor triggering
 The first adjuvant for EAE studies was an oil-in-water
emulsion called IncompleteFreund’s Adjuvant (IFA).
Addition of inactivated dried mycobacteria such as M.
butyricum, M. tuberculosis or Bordetella pertussis.
 This has led to the development of secondary progressive
EAE , models of cortical demyelination and experimental
inflammatory neurodegenerative and spastic diseases.

14. 2. VIRAL
 Several mechanisms have been proposed to explain how viruses can
induce demyelination,and may thus be involved in MS. Damage may
result either from a direct effect on neurons, in which case myelin
damage occurs as a secondary event (the so-called ‘insideout’model
)or from a direct attack on myelin, in which case neurons die due to
the lack of trophic support bymyelin (the so-called ‘outside-in’ model).

15. PROCEDURE :-
Some of the viruses are used to induce the disorder, virus
infection could additionally induceor augment autoimmunity to
myelin and neurons via several pathways.
1.Semliki Forest Virus
2.Japanese Macaque Encephalomyelitis
3. Theiler’s Murine Encephalomyelitis Virus

16. 1.SEMLIKI FOREST VIRUS
 Semliki Forest virus (SFV) does not cause demyelination in
humans.SFV is an enveloped Togavirus, first isolated from
mosquitoes in 1942.The common strains used to induce
myelin damage in mice are the mutant M9 and avirulent A7.
SFV is neuroinvasive.once inoculated peripherally and after
crossing the BBB, the virus infects neurons and
oligodendrocytes. While the M9 strain is highly virulent,
causing death in 10-20% of adult mice or paralysis as a result
of neuronal damage

17. 2. JAPANESE MACAQUE ENCEPHALOMYELITIS
 A spontaneous inflammatory demyelinating disease was
reported in a colony of Japanese macaques in Oregon from
which a gamma-herpes virus was isolated.

18. 3. THEILER’S MURINE ENCEPHALOMYELITIS VIRUS
 Theiler’s Murine Encephalomyelitis Virus, first identified by
Max Theiler, is a natural pathogenof mice,causing paralysis
and encephalomyelitis.Theiler’s Murine Encephalomyelitis
Virus infection induces clinical neurological disease in
immunocompetent mice, along with atrophy of the brain
and spinal cord

19. 3.TOXIN MODELS
 In these models, demyelination is induced after focal
application or systemic administration of the toxin.Agents for
focal demyelination used so far are
•Lysolecithin, also called LPC
•Ethidium bromide (EB)
•Antibodies to oligodendrocyte-related proteins
•Bacterial endotoxins
•6-aminonicotinamide
•Electrolytes or cocktails containing complement and
antibodies
•against galactocerebroside